Neuroscience Antibodies & Protocols (U.S.A. & European edition) 2006

www.abcam.com Catalogue contents agtidx..92 Targetindex Target index ..91 antibodies Neuroscience New Coming soon W ..89 details Contact Ordering andcontactdetails Technical help Neuroendocrinology Neurodegeneration Growth &development Cell adhesionproteins Neuroscience antibodycategories ..5 key Species Abbreviations Neuroscience at ..1 About Abcam Welcome to Abcam Contents ays toorder rdc aahe ud ..65 guide datasheet Product . ..64 at Abcam resources Neuroscience Obesity &Metabolism . .27 Axis HPA . . .26 Gonadotrophic Axis . ..26 Regulation GH Related . .24 Prions . ..22 Parkinson’s Huntington’ ..18 Alzheimer’s ..14 Transduction Signal ..13 Pathway Notch ..12 Neurotrophins ..11 Neurogenesis ..11 Guidance Axonal ..10 Nuclear ..10 Membrane ..8 ECM ..6 Cytoskeletal ersineAwr ..2 Neuroscience Abwire T argets s . . Abcam ..2 Abcam . . Ordering information: 22 28 25 89 www.abcam.com | Abcam’ ..89 ordering for Information Required Neurotransmission Neuronal markers Sensory pathways Applications key ..5 key Applications Abreviews ..3 account an Abcam of Benefits Neuroscience resources: rnpres. . .61 Transporters Secretory V Receptors &Channels . ..53 Tools Pharmacological ..52 Oxide Nitric . . .50 Neuropeptides ..47 Neurotransmitters ..44 Signaling Intracellular ..43 Signaling Calcium ..35 Neuron ..33 Cells Stem Neural Glia ersinesgaigptwydarm ..85 diagrams pathway signaling Neuroscience ..66 tips and protocols Neuroscience V ..63 Touch ..63 Olfaction . ..62Nociception . ..62 Hearing hri xs..30 Thyroid Axis ..30 Vasopressin & Oxytocin h em. ..2 team The ision ..31 s termsandconditions . SM and AbcamAbpoints sce ..59 esicles Tech support: . www.abcam.com/neuroscience . SM www.abcam.com/technical ..3 90 53 63 Welcome to Abcam About Abcam

Abcam, located in Cambridge, U.K. and Cambridge, MA, U.S.A., was founded in 1998 by Dr. Jonathan Milner, then a researcher at Cambridge University, U.K.:

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Neuroscience at Abcam

Our range of over 2,000 Neuroscience primary antibodies has a breadth and depth that is hard to beat. Further more, we collaborate with leading academics to make the Abcam range one of the best in the world!

The Neuroscience Abwire Our Neuroscience Abwire web page is full of exclusive articles, product browsing tools, conference and publication information and specialist protocols. Visit today for cup-to-date information in this rapidly progressing field.

Look out for new Neuronal Marker IHC Galleries, Neurodegeneration and Neurotransmission signaling pathway diagrams and Neuroscience Product Locator tools (to name but a few).

Visit: www.abcam.com/neuroscience for more.

The Neuroscience team Abcam has a specialized team working on sourcing and testing the best Neuroscience antibodies available. Our background in the neurosciences is very strong to better serve our customers. Members include:

Technical Support Sarah and Bill help customers Sarah Mardle (PhD King’s College, University of London, U.K.) who have enquiries about, or Technical Support Manager (U.K.) problems with, Abcam antibodies using their extensive PhD and postdoctoral laboratory experience.They also:

• Attend conferences to keep up-to-date with antibody Bill Campbell (PhD Brown University, U.S.A.) techniques and research Technical Support Manager (U.S.A.) • Source new protein targets

• Source novel techniques using antibodies

• Build collaborations with academics.

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 2 Neuroscience resources: www.abcam.com/neuroscience Neuroscience at Abcam

The Neuroscience team

Business Development Sabrina Edmonds (PhD University of Cambridge, U.K.) Business Development Manager for Neuroscience W Sabrina is responsible for sourcing new and exciting protein targets to generate antibodies against by elcome to attending conferences, forging collaborations with academics and by reading current literature. She is the main contact for academics who are interested in licensing their neuroscience antibodies for sale through Abcam.

Marketing Abcam Rhian Hayward (PhD University of Oxford, U.K.) Neuroscience Marketing Manager

Rhian is responsible for marketing Abcam’s Neuroscience range to our customers worldwide. She ensures that Abcam supports many Neuroscience community activities through conference sponsorships, informative marketing materials e.g. signalling pathways (see page 83), the Neuroscience Abwire and more.

Suggestions or comments: Email the team at [email protected]

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AbreviewTM Action Points Description

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Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 4 Neuroscience resources: www.abcam.com/neuroscience Abbreviations Species key

Key Species Key Species Key Species

Many wide range of species Fe Ferret Pi Pig Fi Fish P Plants At Arabidopsis thaliana Gt Goat Q Quail Am Amphibian G Goose Rb Rabbit Av Avian Gp Guinea pig Rt Rat Ba Baboon Ha Hamster Re Reptiles Bu Burkholderia spp. H Horse Ro Rodents Ca Cat Hu Human Sc Sacchromyces cerevisiae Ce Caenorhabditis elegans In Insect S Salamander Ch Chicken K Kangaroo Sha Shark C Cow Li Lizard Sh Sheep De Deer Mm Mouse Sn Snake D Dog Ma Mammals X Xenopus laevis Abbreviations Dm Drosophila melanogaster Mi Mink Y all Yeast El Elk Mk Monkey Ec Escherichia coli Op Opossum

Applications key

Key Applications Key Applications

AF Acetone fixed IHC-P Immunohistochemistry (Formalin-fixed Agg Agglutination paraffin-embedded sections) AI Adhesion inhibition IM Immunomicroscopy AP Affinity purification Inhib Inhibition assay BL Blocking IP Immunoprecipitation CCIe Counter current IRMA Immunoradiometric assay immunoelectrophoresis ISH In situ hybridization ChIP Chromatin immunoprecipitation Mct Microcytotoxicity testing Conjugation Conjugation Neut Neutralizing DID Double immunodiffusion P Purification Dot Dot blot PA Prothrombin assay EC Experimental control PCC Primary cell culture EIA Enzyme immunoassay PCR Polymerase chain reaction ELISA ELISA RIA Radioimmunoassay EM Electron microscopy RID Radial immunodiffusion EMSA Electrophoretic mobility shift assay RIe Rocket immunoelectrophoresis FACS Fluorescence activated cell sorting RImm Radial immunodiffusion FC Flow cytometry RipA Radioimmunoprecipitation assay FuncS Functional studies SB Southern blot GSA Gel supershift assays TCC Transfected cell culture H Histology TLC Thin layer chromatography I-ELISA Indirect ELISA WB Western blotting IA Immunoassay IB Immunoblotting Other ICC Immunocytochemistry Fast Track Application and species data pending ID Immunodiffusion Pep. avail. Peptide available Ie Immunoelectrophoresis IF Immunofluorescence IHC Immunohistochemistry IHC-F Immunohistochemistry (Formalin fixed sections) IHC-Fr Immunohistochemistry (Frozen sections)

Many of the images in this catalog have been supplied by Abcam’s collaborators in the Neuroscience field. We wish to thank all these contributors and we look forward to working with you in the future!

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Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Intermediate Filaments

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

GARP (truncated) P WB Rb Rt, B ab15094 GFAP P IF, IHC-Fr Ch Hu, Mm, Rt, F, C ab4674 GFAP P IF, IHC-F, IHC-P, IP, PCC, WB Rb F, B, Hu, Mm, Pi, Rt ab7260 GFAP P IF, IHC-F, IHC-Fr, IHC-P Rb Hu, Rt, B ab7779 GFAP [2A5] M IF, IHC-F, IHC-P, PCC, WB Mm Ma ab8136 GFAP [6F2] M IHC-Fr, IHC-P, IM, TEM immunogold labeling, WB Mm Hu ab8975 GFAP [GF-01] M ICC, IHC-Fr, IHC-P, IP, WB Mm Hu, F, Pi Does not react with Mm or Rt ab7806 GFAP (phospho T7) [YC10] M ICC, WB Mm Hu, C, Pi Does not react with Mm or Rt ab12340 Internexin alpha P ICC, IHC-Fr, WB Rb Hu, Mm, Rt, C, F ab22038 Internexin alpha [1D2] M ICC, IHC-P, WB Mm Hu, Rt ab22039 Internexin alpha [2E4] M ICC, IHC-Fr, IHC-P, WB Mm Hu, Rt ab22040 Laminin P ELISA, IF, IHC-P, RIA Rb Hu ab23753 Laminin P ELISA, WB Ch Hu, Mm, Rt ab14055 Laminin P Dot, IF, IHC-F, IHC-P Rb Ma, Am, Av, Re ab11575 Laminin [A5] M ICC, IHC-Fr, WB Rt Hu, Mm, Ba ab2500 Laminin [LAM-89] M IF, IHC-P Mm Hu, F, Rb. Does not react with Mm, Rt, Ch, D, Gp, Li, Pi, Sh or Sn ab11574 Laminin [MEC5] M IHC-Fr Rt Hu, Mm, Rt ab2466 Laminin 2 alpha [4H8-2] M ELISA, ICC, IF, IP, WB Rt Hu, Mm ab11576 Laminin 5 P IHC-Fr, WB Rb Hu ab14509 Laminin B2 gamma 1 [SPM277] M IF, IHC-P, IP, WB Rt Hu, Mm ab17792 Laminin S [C4] M IP, WB Mm Hu, Rt, B, Pi, Gp, Ch ab3289 67kDa Laminin Receptor P ICC, IF, IHC-P, IP, WB Rb Hu, Mm ab711 67kDa Laminin Receptor [MLuC5] M FACS, ICC, IF, IHC-P Mm Hu, Rt ab3099 Nestin P ICC, IF Rb Hu, Ha, Mk

Cell adhesion proteins Does not react with Mm ab5968 Nestin [10C2] M IF, IHC-Fr, IHC-P, WB Mm Hu. Does not react with rodent nestin ab22035 Nestin [2C13A11] M FACS, ICC, IP, WB Mm Hu. Does not react with Rt ab18102 Nestin [2Q178] M EM, ICC, IHC-Fr, IHC-P, WB Mm Mm, Rt Does not react with Hu, F, Mk or Sh ab6142 Nestin [3k1] M ELISA, FACS, ICC, IHC (aldehyde-based fixation), IP, WB Mm Hu ab6320 68kDa Neurofilament [1D2] M ICC, IF, IHC-Fr, IHC-P, WB Mm Hu, Rt ab4571 68kDa Neurofilament [NR4] M IF Mm Hu, Rt, Ch, Pi ab11219 68kDa Neurofilament [SPM204] M IHC-P, WB Mm Hu ab17832 70 kDa Neurofilament Light P IHC-F, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, B, Pi, Av ab9035 70 kDa Neurofilament Light [2F11] M IHC-Fr, IHC-P, WB Mm Hu, Op ab8970 70 kDa Neurofilament Light [DA2] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt, Ch, C, Pi ab4572 160 kDa Neurofilament Medium P IF, IHC-F, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, F, B, Pi, Re, Av ab9034 160 kDa Neurofilament Medium [3H11] M IHC-F, IHC-Fr, IHC-P, WB Mm Hu ab7256 160 kDa Neurofilament Medium [RNF403] M IHC-Fr, WB Mm Hu, Rt ab9271

160 kD Neurofilament Medium antibody [NF-09] (ab7794)

IF - ab7794 at a dilution of 1/1000, staining on rat brain tissue (30µm thick coronal sections) in free floating IHC.

Clonality Applications tested Host Species reactivity Datasheet M WB, ELISA, IHC-P, IHC-Fr, IF, ICC Mm Ma www.abcam.com/ab7794

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 6 Neuroscience resources: www.abcam.com/neuroscience Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Intermediate Filaments

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

200 kDa Neurofilament Heavy P IF, IHC-Fr, IHC-P, WB Rb F, B, Hu, Mm, Pi, Rt ab8135 200 kDa Neurofilament Heavy [NAP4] M ELISA, IHC-F, IHC-Fr, IHC-P, WB Mm Av, Ma ab7257 200 kDa Neurofilament Heavy [RNF402] M IHC-Fr, IHC-P, WB Mm Hu, Rb, Rt, Mm, B, D, Sh, Gp, Ch, Ha, Mk, X ab3966 200 kDa Neurofilament Heavy [SPM203] M IHC-P, WB Mm Hu ab17879 200kDa + 70kDa Neurofilament [2F11] M IF, IHC, IHC-P, IP, WB Mm Hu ab17114 Peripherin P ELISA, ICC, IF, IHC-F, IHC-P, WB Rb Ma ab7258 Peripherin [2Q135] M ICC, WB Mm Hu, Mm, Rt, C, Pi. Does not react with Ch ab17999 Peripherin [7C5] M ICC, IHC-P, WB Mm Ch ab4573 Peripherin [8G2] M ELISA, ICC, IHC-F, IHC-P, WB Mm Ma ab7657 Peripherin variant PER56 P IHC-Fr, IHC-P, WB Rb Hu, Mm ab4655 Peripherin variant PER61 P IHC-Fr, IHC-P, WB Rb Hu, Mm ab4646

200 kD Neurofilament Heavy antibody [NF-01] (ab7795)

IF - ab7795 at a dilution of 1/1000, staining in rat brain tissue (30 µm thick coronal sections) on free floating IHC.

Clonality Applications tested Host Species reactivity Datasheet Cell adhesion proteins M WB, ELISA, IHC-P, IHC-Fr, IF, ICC Mm Ma www.abcam.com/ab7795

Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Microtubules

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/... beta III Tubulin [TU-20] M ICC, IF, IHC-P, IP, WB Mm Ma ab7751 beta III Tubulin [TUJ-1] M ICC, IF, IHC-Fr, WB Mm Hu, Rt ab14545 Doublecortin P IHC-Fr Gp Hu, Mm, Rt ab10293 MAP2 P IF, IHC, WB Ch B, Hu, Mm, Rt ab5392 MAP2 [AP-20] M ICC, WB Mm Mm, Rt, X, B, Q, S ab11268 MAP2 [HM-2] M ICC, IHC, WB Mm Hu, Mm, Rt, B, Ch, Q ab11267 MAP2 [MT-01] M ICC, IF, IP, WB Mm Mm, F, C, Pi ab7756 MAP2a + MAP2b [AP20] M ICC, IF, IHC-P, WB Mm Hu, Rt, Mm, B, Ch, X , Q ab3096 MAP2a + MAP2b [MT-01] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, Mm, F ab8479 MAP5 [3G5] M IF, IHC-P, IHC on primary cultures Mm Hu, Mm, Rt, C. Does not react with Ch ab3095

Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Regulation

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Thrombospondin 1 [CSI 002-65] M ELISA, IHC, IHC-Fr, IP Mm Hu, B ab23420

Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Thin Filaments

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

MYRIP P Fast track Gt Fast track ab10149 Pep. avail.

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Neuroscience > Cell Adhesion Proteins > ECM Proteins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Agrin [AGR 131] M ICC, IHC, IRMA, WB Mm Mm, Rt ab12362 Agrin [Agr 247] M ICC, IF Mm Mm, Rt ab12364 Agrin [Agr 86] M FACS, ICC, WB Mm Mm, Rt ab12361 Amisyn P Fast track Gt Fast track ab5899 Pep. avail. Bestrophin P ICC, IHC, IP, WB Rb Hu, Mm, Rt ab14927 Bestrophin M WB Mm Hu, Pi, Mk ab2182 Drebrin P WB Rb Rt, Huab11068 Drebrin [M2F6] M IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt, Ch, F, Gp, Mk ab12350 Fetuin A P ELISA, WB Rb Hu ab13980 Laminin P ELISA, IF, IHC-P, RIA Rb Hu ab23753 Laminin P ELISA, WB Ch Hu, Mm, Rt ab14055 Laminin P Dot, IF, IHC-F, IHC-P Rb Ma, Am, Av, Re ab11575 Laminin [A5] M ICC, IHC-Fr, WB Rt Hu, Mm, Ba ab2500 Laminin [LAM-89] M IF, IHC-P Mm Hu, F, Rb. Does not react with: Mm, Rt, Ch, D, Gp, Li, Pi, Sh or Sn ab11574 Laminin [MEC5] M IHC-Fr Rt Hu, Mm, Rt ab2466 Laminin 2 alpha [4H8-2] M ELISA, ICC, IF, IP, WB Rt Hu, Mm ab11576 Laminin 5 P IHC-Fr, WB Rb Hu ab14509 Laminin B2 gamma 1 [SPM277] M IF, IHC-P, IP, WB Rt Hu, Mm ab17792 Laminin S [C4] M IP, WB Mm Hu, Rt, B, Pi, Gp, Ch ab3289 67kDa Laminin Receptor P ICC, IF, IHC-P, IP, WB Rb Hu, Mm ab711 67kDa Laminin Receptor [MLuC5] M FACS, ICC, IF, IHC-P Mm Hu, Rt ab3099 Mint3 P WB Rb Rt, Mmab3450 Pep. avail. NCAM [123C3] M IF, IHC-F, IHC-Fr, IHC-P Mm Rt, Hu ab8077 NCAM [B332 (123A8)] M FACS, IHC-Fr, WB Mm Hu ab8596 NCAM [ERIC-1] M ELISA, IHC-Fr, IP, RIA, WB Mm Hu ab6123 NCAM [H28-123] M AP, IHC-Fr, IM, WB Rt Mm ab19782 NCAM [MEM-188] M FACS, IP, WB Mm Hu ab8233 NCAM [RNL-1] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt ab9018 Cell adhesion proteins Nicastrin P WB Rb Hu, Mmab3444 Pep. avail. SIRP [MRC OX-41 ] M FACS, IHC-Fr, IHC-P, WB Mm Rt ab9295 SIRP alpha P IHC-P, WB Rb Hu, Mk. Does not react with Mm ab2971 SIRP alpha P WB Rb Hu, Rt, Mm ab8120 Pep. avail.

67kD Laminin Receptor antibody (ab2508)

IF - staining observed in the hippocampus in the cytoplasm of some astrocytes and neurons using ab2508. Recommended dilution of antibody is 1:300-1:1000 with overnight incubation at room temperature. Free floating IHC performed with TSA amplification.

Clonality Applications tested Host Species reactivity Datasheet P IHC-Fr, Fl, IP, WB Rb Hu www.abcam.com/ab2508

Reelin antibody [142] (ab18569)

IHC-P - ab18569 detecting Reelin immunoreactivity in Cajal-Retzius cells of the mouse cortex at postnatal day 0. ab18569 was used at 1/1000 (overnight incubation at 4°C). Immunoreactivity was visualized using ABC amplification.

Clonality Applications tested Host Species reactivity Datasheet M ELISA, ICC, IHC-P, IP Mm Many www.abcam.com/ab18569

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 8 Neuroscience resources: www.abcam.com/neuroscience Neuroscience > Cell Adhesion Proteins > ECM Proteins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Synaptotagmin P ICC, IP, WB Rb Rt, X ab13260 Synaptotagmin P IF, WB Rb Hu, Rt ab10104 Synaptotagmin P ELISA, WB Ch Hu, B, Mm, Pi, Rt ab8037 Synaptotagmin [ASV30] M ICC, IHC, IP, WB Mm Mm, Rt, X, B, Ch, Rb, Sha ab13259 Synaptotagmin [ASV48] M ICC, IHC, IP, WB Mm Rt, B, Mk, Sha, Mm ab12255 SynCAM P WB Rb Rt ab3910 Syntrophin [1351] M IF, IP, WB Mm Hu, Mm, Rt, Am, Ch, D, Fi ab11425 Syntrophin gamma 2 P ELISA, WB Gt Hu ab15713 Pep. avail. Tenascin C P WB Ch Hu ab16290 Tenascin C [MTn-12] M AF, Dot, ELISA, IF, IHC, IHC-F, IHC-Fr, IP, WB Rt Mm ab6346

Reelin antibody [G10] (ab18570)

IHC-P - ab18570 detecting Reelin immunoreactivity in Cajal-Retzius cells of the mouse cortex at postnatal day 0. ab18569 was used at 1/5000 (overnight incubation at 4°C). Immunoreactivity was visualized using ABC amplification. NB: Antigen retrieval performed (microwave in citrate buffer pH 6).

Clonality Applications tested Host Species reactivity Datasheet M WB, IP, ELISA, IHC-Fr, ICC, IHC-P Mm Mm, Rt. Does not react with Hu www.abcam.com/ab18570 Cell adhesion proteins

Neuroscience Abwire at www.abcam.com/ neuroscience

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Neuroscience > Cell Adhesion Proteins > Membrane Proteins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Cathepsin B P ID, WB Sh Hu ab211 Cathepsin B P IHC-Fr, IHC-P Rb Hu ab7670 Cathepsin B [3E1] M IHC-Fr, IHC-P Mm Hu ab7430 Cathepsin D P ELISA, WB Rb Hu ab19555 Cathepsin D P IHC-Fr, IHC-P Rb Hu ab826 Cathepsin D [CTD-19] M ELISA, IHC-F, IHC-P, WB Mm Hu ab6313 Cathepsin D [D101] M IHC-Fr, IHC-P Mm Hu ab7433 Cathepsin G P ELISA, WB Rb Hu ab20725 Cathepsin G P IHC-Fr, IHC-P Rb Hu ab15988 Cathepsin G P WB Sh Hu ab8816 Cathepsin H [1D10] M IHC-Fr, IHC-P, WB Mm Hu ab7432 Cathepsin L P IHC-Fr, IHC-P Sh Hu ab7454 Cathepsin L [33/2] M ELISA, IHC-Fr, WB Mm Hu, Mm, Rt, Mi ab6314 Heparan Sulfate Proteoglycan 2 [CSI 001-74] M AP, ELISA, IHC, IHC-Fr, IP, WB Mm Hu, C ab23418 Heparan Sulfate Proteoglycan 2 [SPM255], prediluted M IHC-P Rt Hu, Mm, B, Pi ab17848 MOSP [CE1] M IF, IHC-Fr Mm Hu, Rt, Mm, F, Ch, Mk, Ca ab3094 Myelin Basic Protein P WB Rb Hu, Mm, Rt ab12355 Myelin Basic Protein P IF, IHC-Fr, IHC-P Rb Hu, Rt ab2404 Myelin Basic Protein [2B5] M ICC, IHC-F, IHC-Fr, IHC-P Mm Hu ab8053 Myelin Basic Protein [12] M AF, ELISA, IF, IHC-Fr, PCC, RIA, WB Rt Hu, B, Gp, Mm, Rb, Rt, Sh ab7349 Myelin Basic Protein [26] M IHC-Fr Mm MBP from most species ab11162 Myelin Basic Protein [B505] M IHC-Fr, IHC-P Mm Hu, Rb, Rt, C, Mk ab8764 Myelin Basic Protein [Clone 2] M ELISA, IHC-Fr Mm Hu, Rt, B, Gp, Rb, Sh ab11159 Myelin PLP [plpc 1] M IF, IHC-Fr, WB Mm Ma ab9311 PMP22 P IHC-P Rb Hu ab15506 PMP22, prediluted P IHC-P Rb Hu ab15507 PMP22 [20-14] M IHC-P Mm Hu ab3278 SORL1 P Fast track Gt Fast track ab22814 Cell adhesion proteins Spinesin P WB Gt Mm ab2245

Proteins Myelin Basic Protein protein (Biotin) ab792

NrCAM antibody (ab24344)

IF - ab24344 (1/300) detects NrCAM on embryonic day 11.5 mouse spinal cord. Floor plate commissurral axons crossing to the contralateral side of the spinal cord are shown here labeled for NrCAM. Mouse spinal cord was 4% PFA-fixed and processed on slide for IF.

Clonality Applications tested Host Species reactivity Datasheet P IF, IHC-F, IHC-P, IHC-FrFl, IP, WB Rb Hu, Mm, Rt www.abcam.com/ab24344

Neuroscience > Cell Adhesion Proteins > Nuclear Proteins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Galectin 3 P IHC, WB Rb Hu ab2473 Galectin 3 [A3A12] M ELISA, FACS, IF, IHC-Fr, IP, WB Mm Hu, Rt, Mm ab2785 Galectin 7 P IP, WB Rb Mm ab10482

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 10 Neuroscience resources: www.abcam.com/neuroscience

Neuroscience > Growth and Development > Axonal Guidance Proteins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Aggrecan ARGxx [BC-3] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, B, F, D, Gp, H, Pi, Rb, Rt, Sh ab3773 AMIGO2 P WB Rb Mm, Rt ab16762 CRMP5 [CR-1] M IHC-Fr, WB Rt Hu, Rt, B ab19378 CRMP5 [CR-3] M IHC-Fr, WB Rt Hu, Mm, Rt, B ab19352 EphA1 P ELISA, WBRb Hu ab5385 EphA1 P I-ELISA, WBGt Hu ab7036 EphA2 P ELISA, WB Rb Ha, Hu, Mm ab5387 EphA2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610 EphA3 P I-ELISA, WBGt Mm ab7038 EphA3 P ELISA, WB Gt Hu. Does not react with Mm ab10611 EphA3 [III-A4] M FACS, IP Mm Hu ab19439 EphA4 P ELISA, WB Rb Hu ab5389 EphA4 P I-ELISA, WB Gt Mm ab7039 EphA4 P I-ELISA, WB Gt Hu ab7041 EphA5 P ELISA, WB Rb Hu, Mm ab5397 EphA5 P ELISA, WB Gt Mm, Rt ab10612 EphA6 P ELISA, WB Gt Mm ab10613

EphA7 P ELISA, WB Rb Hu ab5411 Growth and development EphA7 P ELISA, WB Gt Mm ab10614 EphA8 P ELISA, WB Gt Mm ab10615 EphB1 P ELISA, WB Rb Hu, Mm ab5414 EphB1 P I-ELISA, WBGt Rt ab7042 EphB1 P Dot, IPSh Hu ab10406 EphB2 P ELISA, WB Rb Hu, Mm ab5418 EphB2 P Dot, ELISA Gt Mm ab10616 EphB3 P ELISA, WBGt Mm ab10617 EphB3 P ELISA, WB Gt Hu ab7044 EphB4 P Dot, ELISA, Inhib Gt Mm ab10618 EphB6 P Dot, ELISAGt Mm ab10619 L1CAM [UJ127] M IF, IHC-P, IP, WB Mm Hu ab3200 L1CAM [UJ12711] M IP, WB Mm Hu ab20148 L1CAM [UJ1814] M IP, WB Mm Hu ab20149 Neuropilin 1 P ELISA, WB Gt Hu ab18130 Neuropilin 1 P IHC-P, WB Rb Rt, Mm ab16786 Neuroserpin P IHC, WB Rb Hu ab16171 NRG1 P WB Rb Hu, Mm ab2994 NRG1 [7D5] M IHC-Fr, IHC-P, IP, WB Mm Rt, Mm ab2369 NRP2 P WB Ch Hu ab15874 Reelin [142] M ELISA, ICC, IHC-P, IP, WB Mm Ma, Av ab18569 Reelin [G10] M ELISA, ICC, IHC-Fr, IHC-P, IP, WB Mm Mm, Rt. Does not react with Hu ab18570 Robo1 P ELISA, IF, WB Rb Mm ab7279 Robo4 P WB Rb Mm ab10547 Slit1 P WB Rb Mm, Rt ab10984 Slit2 P Fast track Rb Fast track ab7665

Proteins Robo4 protein ab19112

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Neuroscience > Growth and Development > Neurogenesis

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Axotrophin P WB Ch Hu, Mm, Rt ab13997 Cdk5 P ELISA, WB Rb Hu, Mm, Rt ab19426 Cdk5 [SPM236], prediluted M IHC-P Mm Hu, Mm, Rt ab17875 DYRK1A P IF, IP, WB Sh Hu ab16140 PTPR P WB Rb Hu ab12153 STMN2 P Fast track Gt Fast track ab21190

FOX1A antibody (ab12161)

ICC - ab12161 staining (1/200) in mouse E11 cortical cells cultured for 6 days.

Clonality Applications tested Host Species reactivity Datasheet P WB, IF, ICC, IHC Rb Hu, Mm www.abcam.com/ab12161

Neuroscience > Growth & Development > Neurotrophins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Artemin P WB Rb Hu ab1136 BDNF P ELISA, WB Rb Hu ab9793 BDNF [35928.11] M ELISA, WB Mm Hu ab10505 BDNF (Biotin) P ELISA, WB Rb Hu ab12385 BMP2 P Dot, ELISA, WB Rb Hu, Mm, Rt ab14933 Growth and development BMP2 P ELISA, WB Rb Hu ab17885 BMP2 [65529.111] M I-ELISA, WB Mm Hu ab6285 BMP2 (Biotin) P ELISA, WB Rb Hu ab17597 CNTF P I-ELISA, Neut, WB Gt Rt ab10834 CNTF P ELISA, Neut, WB Rb Rt ab9785 CNTF P ELISA, IHC, WB Ch Hu, Mm, Rt ab17692 CNTF [AS23] M ELISA, IP, WB Mm Hu ab17284 CNTF (Biotin) P ELISA, WB Rb Rt ab12426 GDNF P I-ELISA, Neut, WB Gt Hu ab10835 GDNF P Dot, IHC-Fr, WB Sh Hu ab6206 GDNF P ELISA, IHC, WB Ch Hu, Mm, Rt ab17637 GDNF family Receptor alpha 2 P IHC-Fr, WB Rb Mm, Rt ab6170 GDNF family Receptor alpha 3 P IHC-Fr, WB Rb Mm, Rt ab6171 GDNF family Receptor alpha 3 P WB Rb Hu, Rt, Mm ab8028 Pep. avail. GDNFR alpha P WB Rb Hu, Rt, Mm ab8026 Pep. avail. GFR alpha 2 P WB Rb Hu, Mm, Rt ab19146

BDNF antibody (ab6201)

IHC - immunostaining BDNF-containing hippocampal neurons in coronal rat brain sections (30 microns). ab6201 used at 1/100 dilution.

Clonality Applications tested Host Species reactivity Datasheet P Dot, ELISA, IHC-Fr, Neut, WB Rb Hu, Rt www.abcam.com/ab6201

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

GFR alpha 3 P WB Rb Hu, Mm, Rt ab19135 LANGF Receptor [MC-192] M FACS, IHC-Fr Mm Rt ab6172 Neurotrophin 3 P Dot, ELISA, IHC-Fr, Neut, WB Rb Hu ab6202 Neurotrophin 3 P ELISA, IHC-Fr, IHC-P, WB Gt Hu, Rt ab10334 Neurotrophin 3 (Biotin) P ELISA, WB Gt Hu ab17524 Neurotrophin 4 P ELISA, Neut, WB Gt Hu ab9824 Neurotrophin 4 M ELISA Mm Hu ab9330 Neurotrophin 4 (Biotin) P ELISA, WB Rb Hu ab17888 Neurotrophin 4/5 P Dot, ELISA, IHC-Fr, Neut, WB Rb Hu ab6205 Neurturin P Dot, ELISA, IHC-Fr, WB Sh Hu ab6208 Neurturin P WB Rb Hu ab8061 Neurturin peptide BL ab8393 NGF P Inhib, Dot, ELISA, IHC-Fr, Neut, WB Rb Mm ab6198 NGF [1.A.15] M ELISA Mm Hu ab18167 NGF 2.5S P Dot, Inhib, WB Rb Hu, Mm, Ch ab10515 NGF beta P ELISA, Neut, WB Gt Hu, Mm, Rt ab10837 NGF beta [25623.1] M Dot, ELISA, Inhib Mm ab10513 NRG1 P WB Rb Hu, Mm ab2994 NRG1 [7D5] M IHC-Fr, IHC-P, IP, WB Mm Rt, Mm ab2369 p75 NGF Receptor P Dot, FACS, IF, IHC-Fr, IHC-P, IP, WB Rb Mm, Rt ab8874 Growth and development p75 NGF Receptor [12A8] M Dot, ICC, IHC-Fr, IP, WB Rt Mm ab8876 p75 NGF Receptor [ME20.4] M EM, ELISA, FACS, ICC, IF, Mm Hu, F, Pi . IHC-P, Inhib, IP, RIA, WB Does not react with Mm, Rt or Ch ab10495 p75 NGF Receptor [NGFR5] M FACS, IF, IHC-P Mm Hu, Ba, F, Ca, Mk, Rb Does not react with Mm or Rt ab3125 Pleiotrophin P ELISA, WB Rb Hu ab14025 Pleiotrophin P ELISA, WB Gt Hu ab10849 SPON1 P WB Ch Hu, Mm, Rt, C ab14271 TrkA (phospho Y490) P ICC, IP, WB Rb Hu, Mm, Rt ab1445 TrkA/B [1.A.37] M IP, WB Mm Hu ab18137 TrkB P IHC-Fr Rb Rt ab6180 TrkB P I-ELISA, WB Gt Hu ab10841

Proteins BDNF protein ab9794 CNTF protein ab9786 GDNF protein ab9790 Neurotrophin 3 protein ab9792 Neurotrophin 4 protein ab9825 Neurturin protein ab9840 NGF beta protein ab9796

TrkA antibody (ab8871)

IF - ab8871 immunostaining in terminals of primary afferent fibers (fibers in the top part of the figure) of the dorsal root ganglion. Free floating IHC performed on 30um cryostat sections. Primary antibody ab8871 was used at 1/3000 and incubated overnight at room temperature.

Clonality Applications tested Host Species reactivity Datasheet P IHC-Fr, IHC-FrFl, IF Rb Rt www.abcam.com/ab8871

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Neuroscience > Growth and Development > Notch Pathway

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

ADAM10 P WB Rb Hu ab10956 APH1a P WB Rb Rt, Hu, Mm ab12104 Pep. avail. APH1a P IF, IP, WB Rb Hu, Mm ab19390 DLL4 P ELISA, IHC-F, IHC-P, WB Rb Hu, Mm ab7280 Jagged 1 P ELISA, FACS, Neut, WB Gt Hu, Rt ab10580 Jagged 2 P Fast track Rb Fast track ab10556 LNX P Fast track Gt Fast track ab4538 MAML1 P Fast track Gt Fast track ab17019 Pep. avail. Notch1 [A6] M FACS, IHC-P, WB Mm Hu, Mm ab3294 Notch1 (activated) P Co-IP, ICC, IF, IP, WB Rb Hu, Mm ab8925 Pep. avail. Notch2 (activated) P Fast track Rb Fast track ab8926 Pep. avail. Notch1 intra P WB Rb Hu. Does not react with Mm ab8387 Pep. avail. Notch1 [SPM206], prediluted M IHC-P Mm Hu, Mm ab17843 Notch1 intra + Notch2 intra P Fast track Rb Fast track ab8928 Pep. avail. Notch 2 intra P WB Rb Hu ab8927 Pep. avail. NUMB P IF, IHC-P, IM, WB Gt Hu, Mm ab4147 Pep. avail. NUMB P IP, WB Rb Hu, Mm ab14140 Pep. avail. NUMBL P Fast track Gt Fast track ab5641 Pep. avail. PEN2 P IP, WB Rb Hu ab16555 Presenilin 1 P ELISA, IHC-Fr, IHC-P, WB Gt Hu ab7297 Presenilin 1, prediluted P IHC-P Rb Hu ab15508 Presenilin 1 [APS 11] M ELISA, IF, IHC-P, WB Mm Hu, Mm, Rt ab15456 Presenilin 1 [APS 18] M ELISA, IF, WB Mm Hu, Mm, Rt ab15458 Presenilin 2 P ELISA, IHC-Fr, IHC-P Gt Hu ab11148 Presenilin 2 P ELISA, IHC, WB Ch Hu, Mm, Rt ab18041 Presenilin 2 [198C67921] M WB Mm Hu ab13926 Presenilin 2 [APS 26] M ELISA, IF, WB Mm Hu, Mm ab15549 SIAH Interacting Protein P Fast track Gt Fast track ab4276 Pep. avail. SIAH1 P WB Gt Hu ab2237 Pep. avail. Growth and development

Neuroscience > Growth and Development > Signal Transduction

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

14-3-3 P IP, WB Rb Hu, Mm, Rt ab9063 14-3-3 P IHC-P, IP, WB Rb Hu, Mm, Rt ab6081 14-3-3 [2Q248] M WB Mm Rt ab14121 14-3-3 [3F7] M IP, WB Mm Hu ab14108 14-3-3 beta P IHC-P, IP, WB Rb Hu, Mm, Rt ab15260 14-3-3 beta P WB Rb Hu, Mm, Rt ab16730 14-3-3 beta [60C10] M ELISA, WB Mm Hu, Mm, Rt ab16859 14-3-3 beta, prediluted P IHC-P Rb Hu, Mm, Rt ab15262 14-3-3 beta + epsilon + zeta [3C8] M WB Mm Hu, Mm, Rt ab12346 14-3-3 beta + zeta P WB Rb Ma, Dm ab14113 14-3-3 beta + zeta [22-IID8B] M ICC, IHC, WB Mm Hu, Mm, Rt, B, D, Mk, Pi, Rb, Sh ab12347 14-3-3 eta + gamma + zeta P ELISA, WB Rb Many ab14124 14-3-3 gamma (acetyl V1) [KC21] M WB Mm Hu ab17005 14-3-3 gamma [3F8] M IP, WB Mm Hu ab14117 14-3-3 sigma [1N6] M IF, IHC-P, IP, WB Mm Hu ab14123 14-3-3 tau [3B9] M IP, WB Mm Hu, Mm, Rt, B ab10439 14-3-3 Binding Motif (phospho S) P ELISA, IHC-P, IP, WB Rb Many ab14127 14-3-3 Binding Motif (phospho S) [3i2] M ELISA, IP, WB Mm Many ab14126 Agrin [AGR 131] M ICC, IHC, IRMA, WB Mm Mm, Rt ab12362 Agrin [Agr 247] M ICC, IF Mm Mm, Rt ab12364 Agrin [Agr 86] M FACS, ICC, WB Mm Mm, Rt ab12361 AKT P WB Ch Hu, Mm, Rt, X , B ab13990 AKT P ICC, IP, WB Rb Hu, Mm, Rt ab6076

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

AKT (unmodified S473) [BDI111] M ELISA, WB Mm Hu ab19442 AKT (phospho S473) P IF Rb Hu, Mm ab20488 AKT (phospho T308) P IF, WB Rb Hu ab8933 AKT (phospho T34) P ELISA, WB Rb Hu, Mm, Rt, Rb ab23509 AKT peptide BL ab9041 CrkL P WB Gt Hu, Mm ab10907 CSN7b P WB Rb Hu ab11895 Ctip1 [14B5] M ICC, IF, IHC, IP, WB Mm Hu, Mm, Fi ab19487 Ctip1 [15E3AC11] M ICC, IF, IHC, IP, WB Mm Hu, Mm ab18688 Ctip1 [18B12DE6] M ICC, IF, IHC, IP, WB Mm Hu, Mm ab19489

AKT antibody (ab8805)

IF - ab8805 used at 1/80 on cultured neonatal rat cardiomyocytes that express a nuclear-targeted AKT construct. AKT staining (green) and actin filaments (red).

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-P, IF, ICC, IP, Rb Hu www.abcam.com/ab8805 Growth and development Transfected Cell Culture, PCC

AKT (phospho S473) antibody (ab8932)

IF - AKT is phosphorylated on Ser 473 in breast tumor. The staining is much stronger than the weak basal level of phosphorylation in normal breast. The staining is cytoplasmic, as in normal tissue. The phospho Ser 473 AKT antibody (ab8932) is used with no pretreatment at a dilution of 1:100.

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-P, IF Rb Hu www.abcam.com/ab8932

AKT (phospho S473) antibody (ab4802)

IF - detection of AKT phospho S473 at a dilution of 1/200 (24h incubation at RT). Perinuclear staining observed in ventral horn neurons of rat spinal cord (30µm saggital sections).

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-P, IHC-Fr, IF, ICC, IHC Rb Hu, Mm, Rt www.abcam.com/ab4802

AKT (phospho T308) antibody (ab4796)

IHC - ab4796 at a dilution of 1/100 (2 overnight incubations at room temperature; ABC amplification). Cytoplasmic and nuclear staining observed in rat spinal cord motorneurons (ventral horn) 30µm saggital spinal cord sections.

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC Rb Hu, Mm, Rt www.abcam.com/ab4796

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Dab1 P IP, WB Gt Mm ab16674 Dab1 P IF, IHC-Fr, IP, WB Rb Mm ab7522 Dab1 (phospho S491) P WB Rb Hu ab5776 Dab1 (phospho Y198) P WB Rb Hu ab5673 Dab1 (phospho Y220) P WB Rb Hu ab5677 Dispatched P Fast track Gt Fast track ab10142 ELAVL2 + ELAVL3 + ELAVL4 [16A11] M IHC-Fr, WB Mm Hu, Mm, Rt, Fi ab14370 EMX2 P IHC, WB Rb Hu, Mm, Rt ab11849 Fetuin A P ELISA, WB Rb Hu ab13980 FMR1 (Drosophila) [6A15] M ELISA, IF, IP, WB Mm Dm. Does not react with Hu ab10299 FOXG1 P WB Gt Hu ab3394 Pep. avail. FOXG1 P IHC-P, WB Rb Hu ab23470 GCNF P IHC-P Rb Hu ab13007 Gemin 1 [2B1] M ELISA, IF, IP, WB Mm Hu, Mm, X ab5831 Gemin 2 [2E17] M ELISA, IF, IP, WB Mm Hu, X ab6084 Gemin 3 [12H12] M ELISA, IF, IP, WB Mm Hu ab10305 Gli1 P IHC-P, IP, WB Rb Hu, Mm ab7523 Gli2 P Fast track Rb Fast track ab7181 Gli2 P Fast track Rb Fast track ab7195 Gli3 P Fast track Rb Fast track ab6050 LIM Kinase P WB Rb Mm ab10561 LIS1 P IP, WBRb Hu ab2607 LIS1 P WB Gt Mm ab2656Pep. avail. MAP4K6 P IHC-P Rb Hu ab13140 NCAM [123C3] M IF, IHC-F, IHC-Fr, IHC-P Mm Rt, Hu ab8077 NCAM [B332 (123A8)] M FACS, IHC-Fr, WB Mm Hu ab8596 NCAM [ERIC-1] M ELISA, IHC-Fr, IP, RIA, WB Mm Hu ab6123 NCAM [H28-123] M AP, IHC-Fr, IM, WB Rt Mm ab19782 NCAM [MEM-188] M FACS, IP, WB Mm Hu ab8233

Growth and development NCAM [RNL-1] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt ab9018 Neuro D4 P Fast track Gt Fast track ab3940 Pep. avail. Neuroglobin P ELISA, WB Ch Hu ab14010 NR2E3 P IHC-P Rb Hu ab13081 NRG1 P WB Rb Hu, Mmab2994 NRG1 [7D5] M IHC-Fr, IHC-P, IP, WB Mm Rt, Mm ab2369 PAX2 P IHC-P, WBRb Hu ab23799 PAX6 P WB Rb Rt, Sh ab5790 Pep. avail. PHOX2A P IHC, IP, WB Rb Hu, Mm, Rt ab12046 PHOX2A P Fast track Gt Fast track ab4511 PHOX2B P IHC, IP, WB Rb Rt, Hu, Mm ab12047 PROX1 P ELISA, IHC-Fr Rb Hu, Mm, Rt, Ch, Fi ab11941 RAI3 P ICC, IHC-PRb Hu ab13274 ROR alpha P WB Rb Hu ab16367 ROR alpha P IHC-P Rb Hu ab13109 ROR beta P IHC-P Rb Hu ab13113 SCP1 P IF Rb Hu, Mmab15087 SCP1 P IF, IHC-Fr, IHC-P Rb Mm ab15090

Ctip2 antibody [25B6] (ab18465)

IHC - using ab18465 on adult mouse hippocampus.

Clonality Applications tested Host Species reactivity Datasheet M WB, IP, IHC-P, IHC-Fr, IF, Rt Hu, Mm, Fi www.abcam.com/ab18465 ICC, IHC, ChIP

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

SIRP alpha P IHC-P, WBRb Hu, Mk. Does not react with Mm ab2971 SIRP alpha P WB Rb Hu, Rt, Mm ab8120 Pep. avail. SIRP [MRC OX-41 ] M FACS, IHC-Fr, IHC-P, WB Mm Rt ab9295 Slit1 P WB Rb Mm, Rtab10984 Slit2 P Fast track Rb Fast track ab7665 SOX2 P IF Rb Hu, Mm ab15830 Pep. avail. SOX6 P IHC, IP, WB Rb Hu, Mm, Rt ab12054 SOX8 P IHC, IP, WB Rb Hu, Mm, Rt ab12055 SOX13 P IHC, IP, WB Rb Hu, Mm, Rt ab12060 SOX14 P IHC, IP, WB Rb Hu, Mm, Rt ab12061 SOX22 P IHC, IP, WB Rb Hu, Mm, Rt ab12062 Sprouty 2 P ELISA, IP, WB Rb Hu ab1043 SynCAM P WB Rb Rt ab3910 Tenascin C P WB Ch Hu ab16290 Tenascin C [MTn-12] M AF, Dot, ELISA, IF, IHC, IHC-F, IHC-Fr, IP, WB Rt Mm ab6346 TRAF6BP P WB Rb Hu ab22049 VCAM1 [14C3] M IF, IHC-P Mm Hu, Mm ab11514

VCAM1 [1G11B1] M FACS, IA, ICC, IHC-Fr, IP Mm Hu ab7219 Growth and development VCAM1 [M/K-2] M FuncS, IF, IHC Rt Mm ab19569 VCAM1 [MVCAMA(429)] M FACS Rt Mm ab23807 Zic1 P IHC, IP, WB Rb Hu, Mm, Rt ab12070 Zic2 P IHC, WB Rb Hu, Mm, Rt ab12072

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Neuroscience > Neurodegeneration > Alzheimer’s > Amyloid

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

ADAM10 P WB Rb Hu ab10956 AMPK alpha 1 P IP, WB Rb B, Hu, Rt ab3759 Amyloid A Component [mcl], prediluted M IHC-Fr, IHC-P Mm Hu ab7505 Amyloid beta Precursor Protein P ELISA, IHC-Fr, IHC-P, WB Gt Hu ab17468 Amyloid beta Precursor Protein P ICC, IP, WB Rb Hu, Mk ab12268 Amyloid beta Precursor Protein P ICC, IHC-P, IP, WB Rb Hu, Mk ab12269 Amyloid Precursor Protein P ELISA, IHC-P, WB Gt Hu ab17325 Amyloid Precursor Protein P IHC-P, IP, WB Rb Hu, Mk ab11117 Amyloid Precursor Protein P WB Rb Hu, Mm ab2073 Amyloid Precursor Protein P ELISA, IHC-Fr, IHC-P, WB Gt Hu ab2084 Amyloid Precursor Protein (phospho T668) P ICC, IHC-P, IP, WB Rb Hu, Mm, Rt ab17417 Amyloid Precursor Protein [3E9] M IHC-P, WB Mm Hu ab12338 Amyloid Precursor Protein Frameshift Mutant P ELISA, IHC-P, WB Gt Hu ab2085 Amyloid Precursor Protein peptide BL ab7874 APH1a P IF, IP, WB Rb Hu, Mm ab19390 APH1a P WB Rb Rt, Hu, Mm ab12104 Pep. avail. beta Amyloid 1-40 P ELISA, IHC-Fr, RIA Rb Hu ab16218 beta Amyloid 1-40 P IP, WB Rb Hu, Mm ab16622 beta Amyloid 1-40 P ELISA, IHC-P, WB Rb Hu ab10147 beta Amyloid 1-40 [3H2] M ELISA, IF, IHC-P, WB Mm Hu ab17331 beta Amyloid 1-40 [BAM-10] M IHC-Fr, IHC-P Mm Hu ab7501 beta Amyloid 1-40 [BDI350] M ELISA, ICC, WB Mm Hu ab20068 beta Amyloid 1-42 P ELISA, IHC-P, WB Rb Hu ab10148 beta Amyloid 17-42 P ELISA Rb Hu ab17327 beta Amyloid P ELISA, IHC, WB Ch Hu ab17472 beta Amyloid P IHC Rb Ro ab14220 beta Amyloid P ELISA, WB Rb Hu, Mm ab2539 beta Amyloid [2C8] M IHC, WB Mm Hu, Mm ab12349 beta Amyloid [4G8] M ELISA, IHC-P, IP, WB Mm Hu ab1910 Neurodegeneration beta Amyloid peptide BL ab3698 CLACP P WB Rb Hu, Mm ab16763 Factor VIII heavy chain [RFFVIIIC/8] M ELISA, WB Mm Hu, Pi. Does not react with Mm, Rt or D ab6345

Amyloid Precursor Protein antibody [MABDE2B4] (ab12266)

IHC-P- ab12266 detecting APP in Alzheimer’s disease human brain. Plaques are observed in the neuropil. Amyloid deposits stained by DE2B4 (ab12266) can be seen around some blood vessels.

Clonality Applications tested Host Species reactivity Datasheet M WB, IP, IHC-P, ICC, Mm Hu, C, Mk, Pi www.abcam.com/ab12266

beta Amyloid antibody [6E10] (ab10146)

IHC-P- using ab1910 [clone 4G8] and ab10146 [clone 6E10]. Stains paraffin - embedded cerebellum.

Clonality Applications tested Host Species reactivity Datasheet M WB, IP, ELISA, IHC-P Mm Hu www.abcam.com/ab10146

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

FE65 P WB Rb Mm ab5668 Pep. avail. FE65 [4H324] M ICC, IP, WB Mm Hu, Mm, Rt ab17469 Humanin P WB Rb Hu ab16758 Mint1 P WB Rb Rt ab3448 Pep. avail. Mint3 P WB Rb Rt, Mmab3450 Pep. avail. PEN2 P IP, WBRb Hu ab16555 RAGE P ELISA, IHC-PGt Hu ab20558 RAGE P IHC-Fr, WB Rb Rt, Mm ab3611 SHC P IP, WB Rb Hu, Mm, Rt ab15039 SHC (phospho Y239 + Y240) P WB Rb Hu ab4890 Pep. avail. SHC (phospho Y317) P WB Rb Hu, Mm ab15040

Neuroscience > Neurodegeneration > Alzheimer’s > Related targets

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Acrolein P ELISA Rb Acrolein ab15138 Caspase 6 (active) P WB Rb Mm, Rt, Hu ab2326

ADAM17 P Dot, WB Rb Hu, Rt, Mm ab2051 Pep. avail. Neurodegeneration ADAM17 P WB Rb Hu, Rt, Pi ab11547 ADAM17 P WB Gt Hu ab13535Pep. avail. Als2 P Fast track Gt Fast track ab4155 Pep. avail. ALS2CR1 P WB Gt Hu ab4123 Pep. avail. ALS2CR2 P WB Gt Hu ab4122 Pep. avail. ALS2CR2 P WB Rb Hu, Mm, Rt ab17205 ALS2CR3 P Fast track Gt Fast track ab4200 Pep. avail. Apolipoprotein E P ELISA, IHC-P, WB Gt Hu ab7620 Apolipoprotein E P ID, IE, RID, WB Rb Mm ab20874 Apolipoprotein E [E6D7] M ELISA, IHC-Fr, IHC-P, IP, WB Mm Hu ab1907 Calpastatin P ELISA, WBCh Hu ab16423 Calpastatin [2G11D6] M ICC, WB Mm Hu, Rt, C, Pi ab3515 Caspase 6 P WB Rb Hu, Mm, Rt ab2552 Caspase 6 P ELISA, WB Rb Hu, Mm, Rt ab18520 Caspase 6 P WB Rb Hu ab19920 Caspase 6 [3F51] M IHC-P, IP, WB Mm Hu ab17866 Caspase 6 [3F52] M IHC-P, IP, WB Mm Hu ab17822 Caspase 6 [6CSP01 (7165)] M IP Mm Hu ab3245 Caspase 6 [6CSP03] M WB Mm Hu ab4062 CCKAR P WB Rb Hu ab14441 Drebrin P WB Rb Rt, Hu ab11068 Drebrin [M2F6] M IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt, Ch, F, Gp, Mk ab12350 ERAB P ELISA, IF, IHC-P, WB Rb Hu ab17298 ERAB [5F3] M Dot, ELISA, IF, IHC-Fr, IHC-P, WB Mm Hu ab10260 Fyn P ELISA, WBGt Hu ab802Pep. avail.

ApoE antibody [D6E10] (ab1906)

IHC-P - ab1906 reactivity with Apo E of vascular amyloid beta in a Down Syndrome tissue section.

Clonality Applications tested Host Species reactivity Datasheet M WB, IHC-P, IHC-Fr Mm Hu www.abcam.com/ab1906

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Fyn P WB Rb Hu, Mm ab13955 Fyn [1S] M IHC-P Mm Hu, Rt, Mm ab3116 Fyn [FYN-01] M ICC, IP, WB Mm Hu, Mm ab1881 PHF2 P WB Rb Hu ab1138 Synuclein (alpha) P H, ICC, IHC, IHC-Fr, IHC-P, WB Sh Hu ab6162 Synuclein (alpha) P IHC-P Rb Hu, Rt ab15530 Synuclein (alpha) P ICC, WB Gp Hu, Mm, Rt, B ab16784 Synuclein (alpha) (phospho Y125) P Fast track Rb Fast track ab10789 Pep. avail. Synuclein (alpha) [4B12] M Dot, ELISA, IHC-Fr, IHC-P, WB Mm Hu. Does not react with Mm or Rt ab1904 Synuclein (alpha) [4D6] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt ab1903 Synuclein (beta) P ICC, IHC-Fr, IHC-P, WB Rb Hu ab6165 Synuclein (beta) P IHC-Fr, IHC-P Sh Hu ab6163 Synuclein (beta) P WB Ch Hu ab16420 Synuclein (beta) P IHC-P Rb Hu, Rt ab15532 Synuclein (gamma) P IHC, IHC-Fr, IHC-P, WB Rb Hu, Rt ab6169 Synuclein (gamma) P IHC-Fr, IHC-P, WB Rb Hu, Rt ab6176 Synuclein (gamma) P IHC-P, WB Rb Hu, Rt ab15534 Ubiquitin P ELISA, IF, IHC-F, IHC-Fr, IHC-P, IP, WB Rb Hu, Rt, H, Mm, B ab7780 Ubiquitin P ELISA, WBRb Y ab14372 Ubiquitin P WB Ch Hu ab19310 Ubiquitin [10C2-2] M AP, ELISA, IHC-Fr, IHC-P, WB Mm All ubiquitinated proteins ab411 Ubiquitin [1B4-UB] M ELISA, FACS, IHC-Fr, IHC-P, WB Mm Hu, B ab122

Ubiquitin antibody (ab6309)

IF - P23H mutant cells immunostained with ab6309 (1/500 dilution) showing co-localization of mutant aggregated Neurodegeneration protein with ubiquitin.

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-Fr, IA, IF Rb C, Sc www.abcam.com/ab6309

Neuroscience > Neurodegeneration > Alzheimer’s > Proteases

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

BACE1 P IHC-P Gt Hu ab11028 BACE1 P WB Rb Hu, Rt, Mm ab2077 Pep. avail. BACE2 P WB Rb Hu, Rt, Mm ab8025 Pep. avail. BACE peptide BL ab5857 Cathepsin H [1D10] M IHC-Fr, IHC-P, WB Mm Hu ab7432 Nicastrin P WB Rb Hu, Mmab3444 Pep. avail. Presenilin 1 P ELISA, IHC-Fr, IHC-P, WB Gt Hu ab7297 Presenilin 1, prediluted P IHC-P Rb Hu ab15508 Presenilin 1 [APS 11] M ELISA, IF, IHC-P, WB Mm Hu, Mm, Rt ab15456 Presenilin 1 [APS 18] M ELISA, IF, WB Mm Hu, Mm, Rt ab15458 Presenilin 2 P ELISA, IHC, WB Ch Hu, Mm, Rt ab18041 Presenilin 2 P ELISA, IHC-Fr, IHC-P Gt Hu ab11148 Presenilin 2 [198C67921] M WB Mm Hu ab13926 Presenilin 2 [APS 26] M ELISA, IF, WB Mm Hu, Mm, Rt ab15549

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Neuroscience > Neurodegeneration > Alzheimer’s > Tangles & Tau

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

14-3-3 tau [3B9] M IP, WB Mm Hu, Mm, Rt, B ab10439 Casein Kinase 1 delta P ELISA, IHC-P Gt Hu ab10877 ERK1 P IHC-P, IP, WB Rb Hu, Rt, Mm ab7947 Pep. avail. ERK1 P ELISA, IP, WB Rb Hu, Rb, Rt, Mm, C, Pi ab9363 ERK1 (HRP) P ELISA, IP, WB Rb Hu, B, Mm, Pi, Rb, Rt ab9364 ERK1 + ERK2 (phospho T185 + T202) P WB Rb Hu ab4819 Pep. avail. ERK1 + ERK2 (phospho T185 + T202) (FITC) P FACS, IHC-Fr, WB Rb Hu ab4820 ERK1 + ERK2 (phospho T202/183 + Y204/185) P WB Rb Hu, Mm, Rt ab16869 ERK1 + ERK2 (phospho T202 + Y204) P IF Rb Hu, Mm, Rt ab20490 ERK1 + ERK2 P IP Sh Hu, Rt ab16573 ERK1 + ERK2 P WB Rb Hu, Mm, Rt, B ab17942 GSK3 (alpha + beta) (phospho Y216 + Y279) P Dot, ELISA, WB Rb Hu, Mm, Rt ab4797 GSK3 (alpha + beta) P IHC-P, WB Rb Hu, Mm, Rt, Ch, Mk ab15314 GSK3 (alpha + beta) [1H8] M ELISA, WB Mm Hu, Mm, Rt, Am ab17059 Neurofibrillary Tangle P IHC-Fr, IHC-P, WB Rb Hu ab2076 Neurofibrillary Tangle P ELISA, IHC-P, WB Rb Hu ab11093 PEN2 P IP, WB Rb Hu ab16555 Pin1 P WB Rb Hu ab12107 Pep. avail. Prealbumin P DID, Ie, RID, RIe Sh Hu, B, F, Ch, D, Gt, Gp, H, Mm, Pi, Rb, Rt ab9015

Prealbumin P IHC, RID Rb Hu, B, F, D, H ab16006 Neurodegeneration Prealbumin P ELISA Gt Hu ab15007 Tau (phospho S199) P WB Rb Hu ab4749 Pep. avail. Tau (phospho S199 + S202) P WB Rb Hu ab4864 Tau (phospho S202) P IHC-Fr, WB Rb Hu ab4839 Pep. avail. Tau (phospho S208) P WB Rb Hu ab10892 Pep. avail. Tau (phospho S214) P WB Rb Hu ab4846 Pep. avail. Tau (phospho S214) P Fast track Rb Fast track ab10891 Pep. avail. Tau (phospho S356) P WB Rb Hu ab4857 Pep. avail. Tau (phospho S262) P WB Rb Hu ab4856 Pep. avail. Tau (phospho S396) P WB Rb Hu ab4858 Pep. avail. Tau (phospho S400) P WB Rb Hu ab4859 Pep. avail. Tau (phospho S404) P WB Rb Hu ab4860 Pep. avail. Tau (phospho S409) P WB Rb Hu ab4861 Pep. avail. Tau (phospho S422) P WB Rb Hu ab4862 Pep. avail. Tau (phospho T181) P WB Rb Hu ab4748 Pep. avail. Tau (phospho T212) P WB Rb Hu ab4842 Pep. avail. Tau (phospho T217) P WB Rb Hu ab4851 Pep. avail. Tau (phospho T231) P WB Rb Hu ab4855 Pep. avail. Tau P IHC-P, WBGt Hu ab11107 Tau P IHC-F, WBRb Hu ab22690 Tau P IF, IHC-Fr, IHC-P Rb Hu ab8763 Tau [T1] M WB Mm Hu, Mm, Rt, B ab12354 Tau [Tau] M IHC-Fr, IHC-P Mm Ch, B, Hu, Mk. Does not react with Mm or Rt ab8739 Tau [TAU-5] M IHC-Fr, IHC-P, WB Mm Hu ab3931

Tau antibody (ab4841)

IF - Transgenic C. elegans first larval stage nematode expressing human wild type Tau GFP (green) and stained with ab4841 (red). Neurons of the head, dorsa and ventral cords and tail region display phosphorylated Tau.

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-Fr, IHC (no paRtffin) Rb Hu www.abcam.com/ab4841 whole animals

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Ubiquitin P IA, IF, IHC-Fr, WB Rb C, Sc ab6309 Ubiquitin P ELISA, IF, IHC-F, IHC-Fr, IHC-P, IP, WB Rb Hu, Rt, H, Mm, B ab7780 Ubiquitin P ELISA, WBRb Y ab14372 Ubiquitin P WB Ch Hu ab19310 Ubiquitin [10C2-2] M AP, ELISA, IHC-Fr, IHC-P, WB Mm All ubiquitinated proteins ab411 Ubiquitin [1B4-UB] M ELISA, FACS, IHC-Fr, IHC-P, WB Mm Hu, B ab122

Neuroscience > Neurodegeneration > Huntington’s

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

HAP40 P IP, WB Rb Mm ab1139 HAP40 P ELISA, WB Rb Hu ab1400 HIPPI P WB Rb Hu ab5205 Huntingtin [1A771] M WB Mm Many ab13583 Huntingtin [HDA3E10] M IF, IHC-Fr, IP, WB Mm Hu, Rt, Mm ab7668 Huntingtin Interacting Protein HIP1 [4B10] M WB Mm Hu ab5402 Huntingtin Interacting Protein HIP1 [4B107], prediluted M IHC-P Mm Hu. Does not react with Mm ab11841 Huntingtin Interacting Protein 2 P ELISA, WB Gt Hu ab21728

Neuroscience > Neurodegeneration > Parkinson’s > Related targets

Target Clonality Applications tested Host Species reactivity Datasheet Neurodegeneration www.abcam.com/...

ALDH1A1 P WB Gt Hu, Mm. Does not react with Rt ab9883 Pep. avail. APH1a P WB Rb Rt, Hu, Mm ab12104 Pep. avail. APH1a P IF, IP, WB Rb Hu, Mm ab19390 CCKAR P WB Rb Hu ab14441 DORFIN P Fast track Gt Fast track ab3997 Pep. avail. DORFIN P Fast track Rb Fast track ab10889

ALDH1A1 antibody (ab24343)

IF - staining of ALDH1A1-containing neurons in adult mouse brain sections using ab24343.

Clonality Applications tested Host Species reactivity Datasheet P IHC-Fr, IHC-FrFl Rb Mm www.abcam.com/ab24343

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

PGP95 P WB Gt Hu ab11145 PGP95 P IHC-Fr Gp Hu, Mm, Rt ab10410 PGP95 P IHC-P Rb Hu ab15503 PGP95 [13C4 / I3C4] M IF, IHC-F, IHC-Fr, IHC-P Mm Hu, Rt, Gp ab8189 PGP95 [31A3] M ELISA, IHC-P, WB Mm Hu, Rt ab20559 PINK1 P IHC-P, WB Rb Hu, Mm, Rt ab23707 SIAH1 P WB Gt Hu ab2237Pep. avail. Synphilin 1 P IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt ab6179 Synphilin 1 P ELISA, IHC-P, WB Gt Hu ab11106

PGP9.5 antibody (ab10404)

IF - ab10404 at a dilution of 1/1000, detecting PGP9.5 in cortical neurons in rat brain tissue sections prepared for free floating IHC (4% PFA fixed, 30µm thick coronal sections).

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-Fr, IF Rb Hu, Mm, Rt, Pi www.abcam.com/ab10404 Neurodegeneration

Neuroscience > Neurodegeneration > Parkinson’s > Parkin / PARK

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

PARK7 P WB Gt Hu, Mm ab4150 Pep. avail. PARK7 [malphaDJ-1/E219] M ICC, WB Mm Hu, Fi. Does not react with Mm ab11251 Parkin P IHC-Fr, WB Rb Hu, Rt ab5667 Pep. avail. Parkin P Dot, ELISA, IHC-Fr, IHC-P, WB Rb Hu, Rt ab6177 Parkin P ELISA, IHC-P, WB Gt Hu ab7296

Parkin antibody (ab15494)

IHC - ab15494 staining Parkin in Alzheimer’s brain (FFPE-sections).

Clonality Applications tested Host Species reactivity Datasheet P IHC-P Rb Hu, Mm, Rt www.abcam.com/ab15494

Neuroscience > Neurodegeneration > Parkinson’s > Synuclein

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

DORFIN P Fast track Rb Fast track ab10889 DORFIN P Fast track Gt Fast track ab3997 Pep. avail. HMGB1 [4C3] M WB Mm Hu, Mm, Rt, C, Pi ab11354 HMGB1 [HAP465] M ELISA, WB Mm Hu, Mm, Rt, Ch, D ab12029 Synphilin 1 P ELISA, IHC-P, WB Gt Hu ab11106 Synphilin 1 P IHC-Fr, IHC-P, WB Rb Hu ab6179 Synuclein (alpha) P ICC, WB Gp Hu, Mm, Rt, B ab16784

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Synuclein (alpha) P IHC-P Rb Hu, Rt ab15530 Synuclein (alpha) P H, ICC, IHC, IHC-Fr, IHC-P, WB Sh Hu ab6162 Synuclein (alpha) (phospho Y125) P Fast track Rb Fast track ab10789 Pep. avail. Synuclein (alpha) [4B12] M Dot, ELISA, IHC-Fr, IHC-P, WB Mm Hu. Does not react with Mm or Rt ab1904 Synuclein (beta) P IHC-P Rb Hu, Rt ab15532 Synuclein (beta) P WB Ch Hu ab16420 Synuclein (beta) P IHC-Fr, IHC-P Sh Hu ab6163 Synuclein (beta) P ICC, IHC-Fr, IHC-P, WB Rb Hu ab6165 Synuclein (gamma) P IHC, IHC-Fr, IHC-P, WB Rb Hu, Rt ab6169 Synuclein (pan) P IHC-P, WB Rb Hu, Rt ab15534 Synuclein (pan) P IHC-Fr, IHC-P, WB Rb Hu, Rt ab6176 VCP [5] M IHC-Fr, IHC-P, IP, WB Mm Hu, Mm, Rt ab11433

HMGB1 (ab11972)

ICC- labeling of HMGB1 (red) in HeLa cells using ab11972 (1/50). DNA is stained with DAPI (blue).

Clonality Applications tested Host Species reactivity Datasheet P IF, WB Rb Hu www.abcam.com/ab11972

alpha Synuclein [4D6] (ab1903) Neurodegeneration IHC-P - ab1903 stains Lewy Bodies in a substantia nigra neuron.

Clonality Applications tested Host Species reactivity Datasheet M ELISA, WB, IHC-P, IHC-Fr Mm Hu, Mm, Rt www.abcam.com/ab1903

Neuroscience > Neurodegeneration > Prions

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

67kDa Laminin Receptor P ICC, IF, IHC-P, IP, WB Rb Hu, Mm ab711 67kDa Laminin Receptor [MLuC5] M FACS, ICC, IF, IHC-P Mm Hu, Rt ab3099 Fyn P WB Rb Hu, Mmab13955 Fyn P ELISA, WBGt Hu ab802 Pep. avail. Fyn [1S] M IHC-P Mm Hu, Rt, Mm ab3116 Fyn [FYN-01] M ICC, IP, WB Mm Hu, Mm ab1881 Prion protease resistant protein 27-30 P ELISA, IHC-P, WB Gt Hu ab7303 Prion protein PrP P ELISA, WB Mm Hu, B ab22366 Prion protein PrP P ELISA, WB Rb Hu, Mm, B ab703 Prion protein PrP P Dot, ELISA, IHC, WB Gt Hu, B, Sh, Ha ab6664 Prion protein PrP [1E5 / G6] M ELISA, WB Mm Hu, B ab2879 Prion protein PrP [1E5 / G6] M ELISA, WB Mm Hu, B ab3408 Prion protein PrP [3B8 / D5] M ELISA, WB Mm C, Sh ab2881 Prion protein PrP [7D9] M ELISA, IHC, IHC-F, IHC-P, WB Mm Mm, B, De, El, Ha, Hu, Rt ab14219 Prion protein PrP [WD3C7] M ELISA, IP, WB Mm B ab13643

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Proteins Prion protein ab1391 Prion protein ab753

67kD Laminin Receptor antibody [SPM278], prediluted (ab18099)

IHC-P - Human breast carcinoma stained with ab18099.

Clonality Applications tested Host Species reactivity Datasheet M IHC-P Mm Hu www.abcam.com/ab18099

Neuroscience > Neurodegeneration > Related targets

Target Clonality Applications tested Host Species reactivity Datasheet Neurodegeneration www.abcam.com/...

5HT2A Receptor P IHC-FrFl, WBRb Rt ab16028 Pep. avail. Als2 P Fast track Gt Fast track ab4155 Pep. avail. ALS2CR1 P WB Gt Hu ab4123 Pep. avail. ALS2CR2 P WB Gt Hu ab4122 Pep. avail. ALS2CR2 P WB Rb Hu, Mm, Rt ab17205 ALS2CR3 P Fast track Gt Fast track ab4200 Pep. avail. Calpain 6 P ELISA, IHC-Fr, IP, WB Rb Hu, Mm, Rt ab16350 CRMP5 [CR-1] M IHC-Fr, WB Rt Hu, Rt, B ab19378 CRMP5 [CR-3] M IHC-Fr, WB Rt Hu, Mm, Rt, B ab19352 Neuroserpin P IHC, WB Rb Hu ab16171 Ubiquitin P ELISA, WB Rb Y ab14372 Ubiquitin P ELISA, IF, IHC-F, IHC-Fr, IHC-P, IP, WB Rb Hu, Rt, H, Mm, B ab7780 Ubiquitin P IA, IF, IHC-Fr, WB Rb C, Sc ab6309 Ubiquitin P WB Ch Hu ab19310 Ubiquitin [10C2-2] M AP, ELISA, IHC-Fr, IHC-P, WB Mm All ubiquitinated proteins ab411 Ubiquitin [1B4-UB] M ELISA, FACS, IHC-Fr, IHC-P, WB Mm Hu, B ab122

HMG14 antibody (ab5212)

ICC - Labeling of HMGN1 (green; ab5212, 1/500) and monoclonal mouse anti-beta-tubulin-II (red) in fixed primary cultures of rat (e18) hippocampal neurons revealed exclusive nuclear staining corresponding to the known nuclear localization of HMG14.

Clonality Applications tested Host Species reactivity Datasheet P IF, WB Rb Hu, Mm, Rt www.abcam.com/ab5212

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Neuroscience > Neuroendocrinology > GH Regulation

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

ENSA P WB Ch Hu, Mm, Rt, B ab14297 GHRHR P IHC-P Rb Hu ab13342 Somatostatin P IHC-F, IHC-Fr, IHC-P Rb Hu ab7840 Somatostatin 14 P Dot, WB Rb Rt ab8903 Somatostatin 14, prediluted P IHC-P Rb Hu ab15522 Somatostatin 28 P Dot, RIA, WB Rb Rt ab8904 Somatostatin Receptor P IHC-Fr, IHC-P Rb Rt, Mm ab2366 Somatostatin Receptor 1 P FACS, WB Rb Hu, Rt ab5827 Somatostatin Receptor 1 P IP, WB Rb Hu, Mm, Rt ab479 Somatostatin Receptor 2 P IF, WB Rb Hu, Mm, Rt ab9550 Somatostatin Receptor 2 P IF, IHC-P Rb Hu, Rt ab13120 Somatostatin Receptor 4 P ELISA, WB Rb Rt ab18796

Proteins Somatostatin 28 protein ab9606 Somatostatin protein ab9607

Somatostatin 28 antibody (ab22682)

EM - ab22682 (1/2000) in guinea pig pancreas D cells. On grid immunogold procedure and 10nm gold particles.

Clonality Applications tested Host Species reactivity Datasheet P EM, IF, IHC-Fr, IHC-P Rb Hu, Mm, Rt, Ch, Cow, Dog, www.abcam.com/ab22682 Cat, Guinea pig, Pig and Sheep Neuroendocrinology

Neuroscience > Neuroendocrinology > Gonadotrophic axis

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Estradiol P RIA Sh Many ab22441 Estradiol [6E1] M RIA Mm Hu ab20257 Estradiol [B42] M ELISA, IHC-Fr Mm Ma ab8743 Estrogen Receptor alpha M IHC-Fr, WB Rb Hu, B ab8617 Estrogen Receptor alpha P WB Ch Hu ab14020 Estrogen Receptor alpha P IP, WB Mm Hu ab283 Estrogen Receptor alpha P ELISA, IHC-P, WB Rb Hu ab5700 Estrogen Receptor alpha [1D5] M IHC-Fr, IHC-P Mm Hu, Rt ab858 Estrogen Receptor alpha [B301] M IHC-F, IHC-Fr, IHC-P, WB Mm Hu ab8857 Estrogen Receptor alpha [B401 (AER303)] M ELISA, IHC-Fr, IP, WB Mm Hu, B ab7820 Estrogen Receptor alpha [EVG F9] M IHC-P, IP, WB Mm Hu, Rb ab19349 Estrogen Receptor alpha [h-151] M GSA, IHC, IP, WB Mm Hu, Mm, Rt, B ab13538

Estrogen Receptor alpha antibody [33] (ab2746)

ICC - mouse 4TI cells stained with ab2746 (1:100). Blue is DAPI stained nuclei, red is Estrogen Receptor alpha.

Clonality Applications tested Host Species reactivity Datasheet M WB, IP, IHC-P, GSA, FACS, IF Mm Hu, Rt, Mm, C www.abcam.com/ab2746

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Estrogen Receptor alpha (phospho S167) P ELISA, IP, WB Rb Hu ab5701 Estrogen Receptor beta P WB Ch Hu, Mm, Rt ab14021 Estrogen Receptor beta P GSA, IHC-Fr, IHC-P, WB Rb Hu, Rt, Mm, Mk ab3577 Pep. avail. Estrogen Receptor beta P IHC, WB Rb Rt ab5784 Estrogen Receptor beta P IHC-Fr, IHC-P, WB Rb Hu, Rt ab5786 Estrogen Receptor beta [14C8] M FACS, IHC, IHC-P, WB Mm Hu ab288 Estrogen Receptor beta [988] M IP, WB Mm Hu, Mm, Rt ab16813 Progesterone Receptor P IHC-P Rb Hu ab15509 Progesterone Receptor [Alpha PR6] M ICC, IHC-Fr, IP, WB Mm Hu, Rb, Rt, Mm, B, Gp, Ch ab2765 Progesterone Receptor [PGR-1A6] M IHC-Fr, IHC-P Mm Hu ab821 Progesterone Receptor [PR-AT 414] M IHC-Fr, IP, WB Mm Hu, Rb, Rt, Mm, B ab2764 Pep. avail. Progesterone Receptor [SP2] M IHC-P, WB Rb Hu ab16661 Progesterone Receptor (phospho S190) P Dot, WB Rb Hu ab13877 Progesterone Receptor (phospho S190) [1154] M WB Mm Hu ab9895 Progesterone Receptor (phospho S294) P Dot, WB Rb Hu ab13878 Progesterone Receptor (phospho S294) [608] M WB Mm Hu ab9896

Neuroscience > Neuroendocrinology > HPA axis

Target Clonality Applications tested Host Species reactivity Datasheet Neuroendocrinology www.abcam.com/...

ACTH P Dot, IHC-Fr Rb C, Rt ab8906 ACTH P IHC-P Rb Hu ab10335 ACTH P RIA Rb Hu, Rt, Mk ab11116 ACTH P ICC, WB Ch Hu, Mm, Rt ab14064 ACTH [56] M ELISA, ICC, IHC-P Mm Hu, Rt ab21003 ACTH [A2H8] M ELISA, RIA Mm Hu ab11114 ACTH [B427] M IF, IHC-F, IHC-Fr, IHC-P, ICC, WB Mm Hu ab8615 CCKBR P IHC-P Rb Hu ab13173 CCKBR P WB Rb Hu ab14439 Corticoliberin [4H9] M ELISA Mm Hu ab10034 Corticotropin Releasing Factor P AP, ICC, RIA Rb Hu, Rt ab8901 Corticotropin Releasing Factor P IHC-Fr Rb Hu, Rt, Sh ab11133 Corticotropin Releasing Factor P IHC, WB Ch Hu, Mm, Rt ab17475 Dehydroepiandrosterone P RIA Rb Hu ab10999 MC4 Receptor P ICC, IHC-Fr, IHC-P Rb Hu ab13034 MC4 Receptor P IHC-P, WB Rb Hu, Mm, Rt ab24233 Melatonin Receptor 1A P IHC-P Rb Hu ab13035 NMUR1 P IHC-P Rb Hu ab13365 NMUR2 P IHC-P Rb Hu ab13366

Proteins ACTH protein ab9586

Corticotropin Releasing Factor Receptor 2 antibody (ab12964)

IF - ab12964 (1/4000) detecting hypothalamic paraventricular CRF Receptor 2-containing neurons in rat brain.

Clonality Applications tested Host Species reactivity Datasheet P IF, IHC-P Rb Hu, Rt www.abcam.com/ab12964

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Neuroscience > Neuroendocrinology > Obesity and Metabolism

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Adiponectin P IP, WB Rb Mm. Does not react with Hu ab3455 Adiponectin P WB Rb Rt ab5596 Adiponectin P ELISA, WB Rb Hu ab17895 Adiponectin P ELISA, Neut, WB Gt Hu ab18065 Adiponectin [19F1] M ELISA, IF, WB Mm Hu, Mm ab22554 Adiponectin [NeNa] M ELISA, IP, WB Mm Hu. Does not react with Mm ab16086 Adiponectin Receptor 1 P WB Mm Mm, Rt ab21814 Attractin P ELISA, WBRb Mm ab14567 Carboxypeptidase H P ICC, IHC-Fr, IP, WB Rb Hu, Mm, Rt, C, Mk ab11053 CCKAR P WB Rb Hu ab14441 Fatty Acid Synthase P IP, WB Rb Hu, Mm, Pi ab3844 GALR2 P IHC-P Rb Hu ab13179 Gastrin Releasing Peptide P IHC-Fr Rb Pi ab22623 Ghrelin P ELISA Rb Hu ab14481 Ghrelin P ELISA, WB Ch Hu, Mm, Rt ab15860 Ghrelin P WB Rb Hu ab19314 Ghrelin [Ghr2] M ELISA, WB Mm Hu ab13982 Ghrelin [Ghr6] M ELISA, WB Mm Hu. Does not react with Mm ab13984 IL6 P ELISA, WB Ch Hu ab14038 IL6 P WB Rb Pi ab11746 IL6 P ELISA, Neut, WB Gt Rt ab9770 IL6 P ELISA, WBGt Rt ab17494 IL6 P ELISA, Neut, WB Gt Hu ab17506 IL6 [5E1] M ELISA, IHC-Fr, Inhib, WB Mm Mk ab8472 IL6 [AS12] (FITC) M FACS Mm Hu. Does not react with Mm ab16169 Neuroendocrinology IL6 [B-E8 ] M ELISA, IHC-Fr, Neut Mm Hu. Does not react with Mm ab11449 IL6 [MP5-20F3] M EIA, Inhib, Neut Rt Mm. Does not react with Hu or Rt ab7375 IL6 [MP5-32C11] M IHC-Fr, Neut Rt Mk. Does not react with Hu or Rt ab7376

GLP1R antibody (ab13181)

IHC-P - ab13181 staining pancreatic formalin fixed paraffin embedded tissue.

Clonality Applications tested Host Species reactivity Datasheet P IHC-P, WB Rb Hu, Rt, Mm, Hamster www.abcam.com/ab13181

IL6 antibody (ab6672)

IHC - ab6672 staining in mouse collagen-induced arthritic tissue.

Clonality Applications tested Host Species reactivity Datasheet P ELISA, RIA, WB, IP, IHC Rb Hu, Mm, Rt www.abcam.com/ab6672

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 28 Neuroscience resources: www.abcam.com/neuroscience Neuroscience > Neuroendocrinology > Obesity and Metabolism

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

IL6 [MQ2-13A5] (FITC) M FACS Rt Hu ab21513 IL6 [MQ2-13A5] (PE) M FACS Rt Hu ab21514 IL6 [MQ2-13A5] M ELISA, FACS, IHC-F, WB Rt Hu ab21515 Insulin Receptor (phospho Y972) P WB Rb Hu, Mm ab5678 Insulin Receptor P ELISA, WB Rb Hu ab5500 Insulin Receptor [29B4] M IP Mm Hu, Mm, Rt ab16884 LAR P WB Rb Hu ab10557 LAR peptide BL ab22637 Leptin P ICC, IHC-Fr, WB Rb Hu, Rt, Mm, Sh ab3583 Leptin P ELISA, WB Ch Mm, Rt ab17531 Leptin P ELISA, WB Rb Mm, Rt ab17629 Leptin [9C10] (HRP) M ELISA Mm Hu ab6059 Leptin [BDI142] M ELISA, ICC, IF, IHC-Fr Mm Hu ab20115 Leptin [BDI170] M IF, IHC-Fr, WB Mm Hu ab20101 Leptin Receptor P IHC-F, WB Rb Hu, Mm, Rt, D ab5593 MC4 Receptor P ICC, IHC-Fr, IHC-P Rb Hu ab13034 Melatonin Receptor 1A P IHC-P Rb Hu ab13035 Melatonin Receptor 1B P IHC-P Rb Hu ab13358 Melatonin Receptor 1B P IHC-P Rb Hu ab13357 Neuropeptide EI NH2 P IHC-Fr Rb Rt ab11141

Neuropeptide Y P IHC, IHC-F, IHC-Fr, IHC-P Sh Hu, Gp, Rb ab6173 Neuroendocrinology Neuropeptide Y P Dot, IHC-Fr, RIA Rb Hu, Mm, Rt, F, B, Mk, Pi, Rb ab10980 Neuropeptide Y P IHC-Fr Gp Rt ab10341 NMUR1 P IHC-P Rb Hu ab13365 NMUR2 P IHC-P Rb Hu ab13366 Orexin A P ELISA, RIA Rb Hu, Mm, Rt ab10981 Orexin A P ELISA, ICC, IHC-Fr Rb Hu ab16787 Orexin B P ELISA, IHC, WB Rb Mm ab14970 Orexin B P ELISA, RIA Rb Hu, Mm, Rt ab10982 Prohormone Convertase 1 P WB Rb Mm ab3532 Pep. avail. Prohormone Convertase 1 P WB Rb Hu, Mk ab16020 Prohormone Convertase 2 P WB Rb Mm ab3533 Pep. avail. Prohormone Convertase 2 P WB Rb Hu, Mk ab12275 PTPIA2 / PTPRN P WB Rb Hu ab12154 SLC6A4 [4A22] M FACS, IHC-P Mm Hu, Rt ab1125 UCP1 P IHC-P, WB Rb Hu, Mm, Rt ab10983

Proteins Adiponectin protein (His) ab13882 IL6 protein ab9731 IL6 protein ab9771 IL6 protein ab9627 Leptin protein ab646 Leptin protein ab7651 Leptin protein ab13987 Leptin protein ab9827 Leptin protein ab9750 Neuropeptide Y protein ab9599

NPY5R antibody (ab13199)

IHC-P - ab13199 staining Hek-293 cells transiently transfected with recombinant NPY5R.

Clonality Applications tested Host Species reactivity Datasheet P ICC, IHC-P Rb Hu www.abcam.com/ab13199

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Neuroscience > Neuroendocrinology > Oxytocin & Vasopressin

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

AVPR1B P IHC-P Rb Hu ab13147 Oxytocin P IHC-P, RIA Rb Hu, Rt, Mm, Rb, Sh ab2078 Oxytocin P IHC-Fr Rb Rt ab11143 Oxytocin Receptor P IHC-P Rb Hu ab13051

Proteins Oxytocin protein ab9600

Neuroscience > Neuroendocrinology > Thyroid axis

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Parathyroid Hormone P IF, IHC-F, IHC-Fr, IHC-P, IP, WB Rb Hu ab7843 Parathyroid Hormone P RIA Rb Hu ab14093 Parathyroid Hormone P ELISA, RIA Gt Hu ab14163 Parathyroid Hormone P ELISA Rb Rt ab14169 Parathyroid Hormone P RIA Gt Rt ab14170 Parathyroid Hormone [A1/70] M ELISA, IHC-Fr, IHC-P, RIA Mm Hu ab14493 Parathyroid Hormone Receptor 1 P IHC-P Rb Hu ab13078 Parathyroid Hormone Receptor 1 [3D11] M IHC-P, WB Mm Hu ab3271 Parathyroid Hormone Receptor 2 P IHC-P Rb Hu ab13080 Thyroid Hormone Receptor [2103] M IHC-Fr, IHC-P, IP, WB Mm Hu ab1131 Thyroid Hormone Receptor alpha 1 P GSA, ICC, WB Rb Ch, Hu ab5621 Thyroid Hormone Receptor alpha 1+2 P IHC-P Rb Hu, Rt, Pi ab15543 Thyroid Hormone Receptor alpha 1+2 [1718] M IHC-P, WB Mm Hu, Mm, Rt, Pi ab16846 Thyroid Hormone Receptor beta P GSA, ICC, WB Rb Hu, Rt ab5622 Thyroid Hormone Receptor beta P IHC-P Rb Hu, D ab15545 Neuroendocrinology Thyroid Hormone Receptor beta [SPM222] M IHC-P Mm Hu, D ab17898 Thyroid Hormone Receptor beta 1 [J52] M GSA, ICC, IP, WB Mm Hu, Rt ab2744 Thyroid Peroxidase [6H7] M AP Mm Hu ab10246 Thyroid Peroxidase [MoAb47] M IHC-P Mm Hu ab12500 TSH P ELISA Gt Hu ab8576 TSH P IHC-Fr, IHC-PRb Hu ab9382 TSH P RIA Gt Hu ab20085 TSH [QB2/6], prediluted M IHC-Fr, IHC-P Mm Hu ab918 TSH [TSH-51] M ELISA, ICC, RIA Mm Hu ab7906 TSH alpha [ME132 ] M ELISA Mm Hu ab9510 TSH beta P ELISA Gt Hu ab21023 TSH beta P ELISA Gt Hu ab20625 TSH beta [ME-130] M ELISA Mm Hu ab9678 TSH beta [ME-130] M ELISA, IA Mm Hu ab9390 TSH Receptor P IHC-P Rb Hu ab13136 TSH Receptor P IHC-P Rb Hu ab13137 TSH Receptor [28] M ELISA, WB Mm Hu, Pi ab2812 TSH Receptor [A10] M ELISA, IHC-Fr, IP, WB Mm Hu ab6048

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Neuroscience > Neuronal Markers > Glia > Astrocyte

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Astrocyte [1B637] M IHC-FrFl Mm Hu, Rt ab14382 Astrocyte [ASTRO1 (10E4/R5)] M IHC-P Mm Hu ab3268 GFAP P IF, IHC-F, IHC-P, IP, PCC, WB Rb Ma ab7260 GFAP [2A5] M IF, IHC-F, IHC-P, PCC, WB Mm Ma ab8136 GFAP [GF-01] M ICC, IHC-Fr, IHC-P, IP, WB Mm Hu, F, Pi. Does not react with Mm or Rt ab7806 GFAP (phospho T7) [YC10] M ICC, WB Mm Hu, C, Pi. Does not react with Mm or Rt ab12340

GFAP antibody (ab7779)

IHC - ab7779 (1/1000) staining GFAP in astrocytes on coronal rat brain tissue sections.

Clonality Applications tested Host Species reactivity Datasheet P IF, IHC-Fr, IHC-P, IHC-F Rb Hu, Rt, Cow www.abcam.com/ab7779 Neuronal markers

GFAP antibody (ab4674)

IF - ab4674 staining rat astrocytes at a dilution of 1:50 (red). Hoechst dye reveals nuclear DNA in blue.

Clonality Applications tested Host Species reactivity Datasheet P IF, IHC-Fr Ch Hu, Mm, Rt, Ca, C www.abcam.com/ab4674

GFAP antibody [6F2] (ab8975)

EM - GFAP (1/100) immunogold labeling of TEM corticobasal degeneration in human brain tissue showing heavy and highly-specific labeling over a glial process.

Clonality Applications tested Host Species reactivity Datasheet M EM, IHC-P, IHC-Fr, WB Mm Hu www.abcam.com/ab8975

S100 antibody (ab7853)

IHC - ab7853 (1/1000) staining of S100 on 30µm coronal rat brain sections. Image shows neuronal cell body and processes.

Clonality Applications tested Host Species reactivity Datasheet P IF, IHC-P, IHC-F, IHC-Fr Rb Hu, Rt and Cow www.abcam.com/ab7853

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

S100 P IF, IHC-P Rb Hu, Rt, Mm, C, Pi ab868 S100 [4E3 ] M IA, P, WB Mm Hu ab10202 S100 [6G1] (HRP) M ELISA, IA, P, WB Mm Hu ab10203 S100 [8B10] M EIA, ELISA, IF, IHC-P, WB Mm Hu, Rt ab8330 S100 [B321] M IF, IHC, WB Mm Hu, Rt, Mm, C, Pi ab7852 S100 [B321] (FITC) M IF, IHC-Fr, IHC-P Mm Hu, B ab8570 S100A8 + S100A9 [27E10] M FACS, IA, IHC-Fr, IHC-P, IP, WB Mm Hu, Rhesus Mk ab17050

Neuroscience > Neuronal Markers > Glia > Microglia

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

CD11b antibody [44] M AF, FACS, FuncS Mm Hu ab19674 CD11b antibody [44] M AF, FACS, FuncS Mm Hu ab19668 CD11b antibody [ICRF44] M FACS, IF Mm Hu ab11355 CD11b antibody [LT11] M IA Mm Hu ab10324 CD11b antibody [M1/70.15] M FACS Rt Mm ab22371 CD11b antibody [M1/70.15] (PE) M FACS, IHC-P Rt Mm ab24518 CD11b antibody [M1/70.15] M FACS, ICC, IHC-F, IHC-P, P Rt Hu, Mm, Rb ab6332 Pep. avail. CD11b antibody [M1/70] M BL, FACS, IHC-Fr Rt Mm ab19628 CD11b antibody [MEM-174] M FACS, FuncS, IP, WB Mm Hu ab1046 CD11b antibody [OX42] M FACS, IHC-Fr, IHC-P, IP Mm Rt ab19576 CD11b/c equivalent antibody [OX42] M FACS Mm Rt ab21568 CD11b/c equivalent antibody [OX42] M ELISA, FACS, IHC-P Mm Rt ab1211 CD11b/c equivalent antibody [OX42] M FACS, IHC-P Mm Rt ab1212 CD45 antibody P WB Rb Hu ab10558

Neuronal markers CD45 antibody [13/2.3] (Biotin) M FACS, IHC-Fr Rt Mm ab19592 CD45 antibody [GA90] (FITC) M FACS Mm Hu ab1175 CD45 antibody [IBL-3/16] M FACS, IHC-Fr, IP Rt Mm. Does not react with Rt or Hu ab23910 CD45 antibody [K252.1E4] M FACS, IHC-Fr Mm Pi ab23918 CD45 antibody [L12/201] M FACS, IHC-Fr, IP Mm Rb ab19566 CD45 antibody [MEM-28] M FACS, IF, IHC, IHC-P, IP, WB Mm Hu ab8216 CD45 antibody [MRC OX-1] (PE) M FACS Mm Rt ab22269 CD45 antibody [MRC OX-1] (FITC) M FACS Mm Rt ab22349 CD45 antibody [PD7/26/16 + 2B11] M IHC, IHC-Fr, IHC-P, IM Mm Hu ab781 CD45 antibody [YKIX 716.13] M FACS, IP Rt D ab23641 CD45 antibody [YW 62.3] (FITC) M FACS Rt Mm ab22475 CD45 antibody [YW 62.3] M FACS, IHC-Fr, IP Rt Rt ab22479 CD45 peptide M BL ab17550 CD45 peptide M BL ab17553 HLA DR + DP + DQ antibody [CR3/43] M IHC-P Mm Hu ab17101 HLA DR + DP + DQ antibody [WR18] (PE) M FACS Mm Hu ab23901 MRP8 antibody [MRP8 7C12/4] M ELISA, FACS, WB Mm Hu ab20220 pan Macrophage antibody M IHC-Fr, IHC-P Mm Rt ab15637 pan Macrophage antibody [BM8] M FACS, IHC-Fr, IHC-P Rt Hu, Mm ab15694 pan Macrophage antibody [RM-1] M FACS, IHC-Fr Mm Rt ab15615

Neuroscience > Neuronal Markers > Glia > Oligodendrocyte

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

CNPase [11-5B] M Dot, ELISA, ICC, IHC-Fr, Mm B, D, Hu, Mm, Pi, Rb, Rt, Sh. ab6319 Pep. avail. IHC-P, IP, WB Does not react with Ch or Gp MOSP [CE1] M IF, IHC-Fr Mm Hu, Rt, Mm, F, Ch, Mk, Ca ab3094 Pep. avail. Myelin [2B5] M ICC, IHC-F, IHC-Fr, IHC-P Mm Hu ab8053

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Myelin Basic Protein P WB Rb Hu, Mm, Rt ab12355 Myelin Basic Protein P IF, IHC-Fr, IHC-P Rb Hu, Rt ab2404 Myelin Basic Protein [12] M AF, ELISA, IF, IHC-Fr, PCC, RIA, WB Rt Hu, B, Gp, Mm, Rb, Rt, Sh ab7349 Myelin Basic Protein [B505] M IHC-Fr, IHC-P Mm Hu, Rb, Rt, C, Mk ab8764 Myelin Basic Protein [Clone 1 (129-138)] M ELISA, IHC-Fr Mm Hu, Rt, B. Does not react with Gp ab11160 Myelin Basic Protein [Clone 2] M ELISA, IHC-Fr Mm Hu, Rt, B, Gp, Rb, Sh ab11159 Myelin PLP [plpc 1] M IF, IHC-Fr, WB Mm Ma ab9311 NRG1 P WB Rb Hu, Mmab2994 NRG1 [7D5] M IHC-Fr, IHC-P, IP, WB Mm Rt, Mm ab2369 Oct4 P IHC-Fr, IP, WB Rb Hu, Mm, Rt ab18976 Pep. avail. Oligodendrocyte Myelin Glycoprotein [F3-87-8] M IHC-Fr, IP Mm Hu, Rt, D ab24022 Oligodendrocyte Specific Protein OSP P IHC-P, WB Rb Mm, Rt ab7474 PMP22 P IHC-P Rb Hu ab15506 PMP22 [20-14] M IHC-P Mm Hu ab3278 Survivin P WB Rb Hu ab8228 Survivin [321] M IF, IP, WB Mm Hu ab10584 Survivin [6011] M IF, WB Mm Hu, Mm, Rt ab4470 Survivin [8E2] M IF, IHC-P Mm Hu, Rt ab3258 Survivin (phospho T34) P WB Rb Hu ab10720 Survivin peptide BL ab7866 Survivin peptide BL ab7865 Neuronal markers

Proteins Myelin Basic Protein protein (Biotin) ab792

Survivin antibody (ab469)

ICC - HeLa cells in prometaphase, metaphase and anaphase stained with anti-Survivin (green), anti-tubulin (red) and DAPI (blue).

Clonality Applications tested Host Species reactivity Datasheet P WB, IP, IHC-F, IHC-P, H, AF, Rb Ca, D, Hu, Mm www.abcam.com/ab469 IHC-Fr, IF, ICC, IHC

Neuroscience > Neuronal Markers > Neural Stem Cell

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Aggrecan ARGxx [BC-3] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, B, F, D, Gp, H, Pi, Rb, Rt, Sh ab3773 BMP2 P Dot, ELISA, WB Rb Hu, Mm, Rt ab14933 BMP2 P ELISA, WB Rb Hu ab17597 BMP2 [65529.111] M I-ELISA, WB Mm Hu ab6285 CD24 [79] M FACS, IHC Rt Mm ab19582 CD24 [ALB9] M ICC Mm Hu ab19704 CD24 [SN3] M IHC-Fr Mm Hu ab3274 CD132 P ELISA, ICC, IF, WB Rb Hu ab16518 CD133 M ELISA, WB Mm Hu ab5558 CNTF P ELISA, WBRb Rt ab12426 CNTF P I-ELISA, Neut, WB Gt Rt ab10834 Pep. avail. CNTF P ELISA, Neut, WB Rb Rt ab9785 Pep. avail. CNTF P ELISA, IHC, WB Ch Hu, Mm, Rt ab17692 CNTF [AS23] M ELISA, IP, WB Mm Hu ab17284

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

EMX2 P IHC, WB Rb Hu, Mm, Rt ab11849 Integrin beta 1 [4B7] M FACS, H, IA, IF, IHC, IHC-Fr, IHC-P, IP, WB Mm Hu ab35 Pep. avail. Integrin beta 1 [4B7R] M FACS, IF, IHC-P Mm Hu, Pi ab3167 Pep. avail. Integrin beta 1 [HM beta 11] (Biotin) M FACS Ha Mm ab23825 Integrin beta 1 [HM beta 11] M FACS, IP Ha Mm ab23833 Integrin beta 1 [HM beta 11] (FITC) M FACS Ha Mm ab23834 Integrin beta 1 [MEM-101A] (FITC) M FACS Mm Hu, Mm, Pi ab21845 Integrin beta 1 [MEM-101A] M FACS, IP, WB Mm Hu. Does not react with C ab8238 Integrin beta 1 (phospho T788 + T789) P WB Rb Hu ab5189 Nestin [10C2] M IF, IHC-Fr, IHC-P, WB Mm Hu. ab22035 Nestin [2C13A11] M FACS, ICC, IP, WB Mm Hu. Does not react with Rt ab18102 Nestin [2Q178] M EM, ICC, IHC-Fr, IHC-P, WB Mm Mm, Rt. Does not react with Hu, F, Mk or Sh ab6142 Nestin [3k1] M ELISA, FACS, ICC, IHC (aldehyde-based fixation), IP, WB Mm Hu ab6320 PAX2 P IHC-P, WBRb Hu ab23799 SOX2 P IF Rb Hu, Mm ab15830 SOX22 P IHC, IP, WB Rb Hu, Mm, Rt ab12062 Vimentin P IF Ch Hu, Mm, Rt ab16521

Integrin beta 1 (phospho S785) antibody (ab5185)

ICC - ab5185 staining showing integrin 1 beta (1/5000) in E11 cortical cells cultured for 6 days. Co-staining for beta III tubulin (not shown) occurs. Neuronal markers

Clonality Applications tested Host Species reactivity Datasheet P WB, IF, ICC, IHC Rb Hu, Ch, Mm www.abcam.com/ab5185

Nestin antibody (ab5968)

ICC - ab5968 (1/200) detecting Nestin immunoreactivity in human embryonic stem cells differentiated in conditions promoting neuronal differentiation.

Clonality Applications tested Host Species reactivity Datasheet P ICC, IF Rb Hu, Mk. Does not react with Mm www.abcam.com/ab5968

Vimentin antibody (ab7783)

IHC-P - labeling in immunoreactive dermal cells in skin of Balb/c mice using ab7783 in 1% bovine serum albumin for 30 min at 37°C.

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-P, IHC-Fr Rb Hu, Mm, Rt www.abcam.com/ab7783

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Vimentin P ICC, IF, WB Gt Hu ab11256 Vimentin P IF Ch Rt ab24525 Vimentin [SP20] M IHC-P Rb Hu ab16700 Vimentin [V9] M FACS, IHC-F, IHC-Fr, IHC-P, WB Mm Hu, Rt, F, Ch, D, H, Pi. Does not react with Mm ab8069 Vimentin [VI-01] M ELISA, ICC, WB Mm Ma ab7752 Vimentin [VI-10] M ICC, IP, WB Mm Hu, Mm, Rt, Ch, Pi ab20346 Vimentin (phospho S55) [4A4] M ELISA, ICC, IHC, WB Mm Hu, Mm, Rt ab22651 von Willebrand Factor P IF, IHC-P, IHC-Fr, IM Rb Gp, Hu, Mm, Rt. Does not react with Ch ab6994

Proteins CNTF protein ab9786

Vimentin antibody [RV202] (ab8978)

IHC-Fr - vimentin staining of a tonsilar lymphoma with ab8978. The epithelium (on the left) is negative.

Clonality Applications tested Host Species reactivity Datasheet

M WB, IHC-Fr, ICC, FACS, IF Mm Hu, Ch, D, Gt, Ha, Mk, www.abcam.com/ab8978 Neuronal markers Mm, Rt, C

Neuroscience > Neuronal Markers > Neuron > Dendrite

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Drebrin P WB Rb Rt, Hu ab11068 Drebrin [M2F6] M IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt, Ch, F, Gp, Mk ab12350 MAP2 [AP-20] M ICC, WB Mm Mm, Rt, X, B, Q, S ab11268 MAP2 [HM-2] M ICC, IHC, WB Mm Hu, Mm, Rt, B, Ch, Q ab11267

MAP2 antibody (ab5392)

IF - ab5392 at a dilution of 1/10000, staining MAP 2 (green) in tissue cultured rat cortical neurons. Nuclei are stained blue with DAPI.

Clonality Applications tested Host Species reactivity Datasheet P WB, IF, IHC-FrFl Ch C, Hu, Mm, Rt www.abcam.com/ab5392

MAP2 antibody [MT-01] (ab7756)

IHC - ab7756 at a dilution of 1/1000, staining MAP-2 on rat brain tissue (30µm thick coronal sections). Labeling of MAP-2 on cortical neuronal processes/axons.

Clonality Applications tested Host Species reactivity Datasheet M WB, IP, IF, ICC Mm Mm, Ca, C, Pi www.abcam.com/ab7756

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

MAP2a + MAP2b [AP20] M ICC, IF, IHC-P, WB Mm Hu, Rt, Mm, B, Ch, X , Q ab3096 MAP2a + MAP2b [MT-01] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, Mm, F ab8479 MAP5 [3G5] M IF, IHC-P, IHC on primary cultures Mm Hu, Mm, Rt, C. Does not react with Ch ab3095 neuron specific beta III Tubulin [TU-20] M ICC, IF, IHC-P, IP, WB Mm Ma ab7751 neuron specific beta III Tubulin [TUJ-1] M ICC, IF, IHC-Fr, WB Mm Hu, Rt ab14545 SAP102 P WB Gt Mm ab12086 SAP102 P IHC-Fr, WBRb Rt. Does not react with Mm ab3438

Neuroscience > Neuronal Markers > Neuron > Growth Cone

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Agrin [AGR 131] M ICC, IHC, IRMA, WB Mm Mm, Rt ab12362 Agrin [Agr 247] M ICC, IF Mm Mm, Rt ab12364 Agrin [Agr 86] M FACS, ICC, WB Mm Mm, Rt ab12361 Axonal Growth Cones [SPM163], prediluted M IHC-P Mm Hu ab17804 BAI1 associated protein 2 isoform 3 P WB Gt Hu ab791 BAIAP2 P WB Gt Hu ab15697 CRMP5 [CR-1] M IHC-Fr, WB Rt Hu, Rt, B ab19378 CRMP5 [CR-3] M IHC-Fr, WB Rt Hu, Mm, Rt, B ab19352 Doublecortin P IHC-Fr Gp Hu, Mm, Rt ab10293 Drebrin P WB Rb Rt, Hu ab11068 Drebrin [M2F6] M IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt, Ch, F, Gp, Mk ab12350

Neuronal markers EphA1 P I-ELISA, WB Gt Hu ab7036 EphA1 P ELISA, WBRb Hu ab5385 EphA2 P ELISA, WB Rb Ha, Hu, Mm ab5387 EphA3 P I-ELISA, WBGt Mm ab7038 EphA3 [III-A4] M FACS, IP Mm Hu ab19439 EphA4 P I-ELISA, WB Gt Mm ab7039 EphA4 P ELISA, WBRb Hu ab5389 EphA5 P ELISA, WB Rb Hu, Mm ab5397 EphA5 P ELISA, WB Gt Mm, Rt ab10612 EphA6 P ELISA, WB Gt Mm ab10613 EphA7 P ELISA, WB Rb Hu ab5411 EphA7 P ELISA, WBGt Mm ab10614 EphA8 P ELISA, WBGt Mm ab10615 EphB1 P I-ELISA, WBGt Rt ab7042 EphB1 P ELISA, WB Rb Hu, Mm ab5414 EphB1 P Dot, IPSh Hu ab10406 EphB2 P ELISA, WB Rb Hu, Mm ab5418 EphB2 P Dot, ELISAGt Mm ab10616 EphB3 P ELISA, WBGt Mm ab10617 EphB4 P Dot, ELISA, Inhib Gt Mm ab10618 EphB6 P Dot, ELISA Gt Mm ab10619 Ephrin A2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610 Ephrin A3 P ELISA, WB Gt Hu. Does not react with Mm ab10611 Ephrin A4 P I-ELISA, WB Gt Hu ab7041 Ephrin B3 P ELISA, WB Gt Hu ab7044 GAP43 P H, ICC, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch. Does not react with X ab7462 GAP43 P ICC, IHC-Fr, IP, WB Rb Hu, Mm, Rt, F, Fi, Mk ab12274 GAP43 (phospho S41) P WB Rb Rt ab3909 Growth Cone [2G13] M IHC-Fr, IHC-P, WB Mm Rt, Mm, Ch ab7762 NCAM [123C3] M IF, IHC-F, IHC-Fr, IHC-P Mm Rt, Hu ab8077

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

NCAM [B332 (123A8)] M FACS, IHC-Fr, WB Mm Hu ab8596 NCAM [ERIC-1] M ELISA, IHC-Fr, IP, RIA, WB Mm Hu ab6123 NCAM [H28-123] M AP, IHC-Fr, IM, WB Rt Mm ab19782 NCAM [MEM-188] M FACS, IP, WB Mm Hu ab8233 NCAM [RNL-1] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt ab9018 Neuroserpin P IHC, WBRb Hu ab16171 NRP2 P WB Ch Hu ab15874 p35 P WB Rb Hu ab10570 p35 [ES19] M IP Mm Hu ab3222 p35 nck5a [SPM430] M IP Mm Hu ab19933 Robo4 P WB Rb Hu, Rb, Sh ab10547

Proteins Robo4 protein ab19112

Neuroscience > Neuronal Markers > Neuron > Soma

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

70 kDa Neurofilament Light P IHC-F, IHC-Fr, IHC-P, WB Rb Ma, Av ab9035 Neuronal markers 70 kDa Neurofilament Light [2F11] M IHC-Fr, IHC-P, WB Mm Hu, Op ab8970 70 kDa Neurofilament Light [DA2] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt, Ch, C, Pi ab4572 160 kDa Neurofilament Medium P IF, IHC-F, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, F, B, Pi, Re, Av ab9034 160 kDa Neurofilament Medium [3H11] M IHC-F, IHC-Fr, IHC-P, WB Mm Hu ab7256 160 kDa Neurofilament Medium [NF-09] M ELISA, ICC, IF, IHC-Fr, IHC-P, WB Mm Ma ab7794 160 kDa Neurofilament Medium [RNF403] M IHC-Fr, WB Mm Hu, Rt ab9271 200 kDa Neurofilament Heavy P IF, IHC-Fr, IHC-P, WB Rb F, B, Hu, Mm, Pi, Rt ab8135 200 kDa Neurofilament Heavy [NAP4] M ELISA, IHC-F, IHC-Fr, IHC-P, WB Mm Ma ab7257 200 kDa Neurofilament Heavy [RNF402] M IHC-Fr, IHC-P, WB Mm Hu, Rb, Rt, Mm, B, D, Sh, Gp, Ch, Ha, Mk, X ab3966 200 kDa Neurofilament Heavy [SPM203] M IHC-P, WB Mm Hu ab17879 200 kDa + 70kDa Neurofilament [2F11] M IF, IHC, IHC-P, IP, WB Mm Hu ab17114 ALK c [ALKc] M IHC-Fr, IHC-P Mm Hu ab650

BrdU (ab1893)

IF - Tissue culture cells were labelled prior to transplantation and then identified in in vivo tissue using ab1893 (10 ug/mL) with a TRITC conjugated secondary antibody.

Clonality Applications tested Host Species reactivity Datasheet P ELISA, IHC-P, IHC-Fr, IP Sh Many www.abcam.com/ab1893

BrdU [BU1/75 (ICR1)] (ab6326)

Astrocytes from adult (in the lesioned cerebellum) were stained with ab6326. (Green: BrdU; Red: Vimentin).

Clonality Applications tested Host Species reactivity Datasheet M FACS, IHC-P, IHC-Fr Mm Many www.abcam.com/ab6326

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Ataxin 7 P WB Rb Hu ab11434 Choline Acetyltransferase P IHC-F, WB Sh Hu, Rt, Rb, Gp, Pi ab18736 Choline Acetyltransferase P ELISA, IHC Rb Mm, Pi, Rt ab6168 CNS gp130 [F3-87-8] M IHC-Fr Mm Mm ab9310 Pep. avail. Coilin [Pdelta] M ICC, WB Mm Hu. Does not react with Mm ab11822 Doublecortin P IHC-Fr Gp Hu, Mm, Rt ab10293 ELAVL2 + ELAVL3 + ELAVL4 [16A11] M IHC-Fr, WB Mm Hu, Mm, Rt, Fi ab14370 ELAVL4 [16C12] M IHC-Fr, IHC-P, WB Mm Hu ab14369 GAP43 P H, ICC, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch. Does not react with X ab7462 GAP43 P ICC, IHC-Fr, IP, WB Rb Hu, Mm, Rt, F, Fi, Mk ab12274 GAP43 (phospho S41) P WB Rb Rt ab3909 GARP (truncated) P WB Rb Rt, B ab15094 HB9/HLXB9 P EMSA, IHC, IP, WB Rb Hu ab12028 MAP2 P IF, IHC, WB Ch B, Hu, Mm, Rt ab5392 MAP2 [AP20] M WB Mm Hu, Mm, Rt, C, Ch ab12342 MAP2 [HM-2] M ICC, IHC, WB Mm Hu, Mm, Rt, B, Ch, Q ab11267 MAP2 [MT-01] M ICC, IF, IP, WB Mm Mm, F, C, Pi ab7756 MAP2a + MAP2b [AP20] M ICC, IF, IHC-P, WB Mm Hu, Rt, Mm, B, Ch, X , Q ab3096 Pep. avail. MAP2a + MAP2b [MT-01] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, Mm, F ab8479 Necdin P WB Rb Hu, Mm, Rt ab18554 Nestin P ICC, IF Rb Does not react with Mm ab5968 Nestin [10C2] M IF, IHC-Fr, IHC-P, WB Mm Hu ab22035 Nestin [2C13A11] M FACS, ICC, IP, WB Mm Hu. Does not react with Rt ab18102 Nestin [2Q178] M EM, ICC, IHC-Fr, IHC-P, WB Mm Mm, Rt. Does not react with Hu, F, Mk or Sh ab6142

Neuronal markers Nestin [3k1] M ELISA, FACS, ICC, IHC (aldehyde-based fixation), IP, WB Mm Hu ab6320 neuron specific beta III Tubulin [TU-20] M ICC, IF, IHC-P, IP, WB Mm Ma, Fi ab7751 neuron specific beta III Tubulin [TUJ-1] M ICC, IF, IHC-Fr, WB Mm Hu, Rt ab14545

c-fos antibody (ab5794)

ICC - ab5794 detecting nuclear staining of c-Fos (green) in adult dorsal root ganglion cells cultured for 2 days. ab5794 was used at 1/100 (incubated for 2 hours at room temperature).

Clonality Applications tested Host Species reactivity Datasheet P ICC Rb Mm www.abcam.com/ab5794

MAP5 antibody [3G5] (ab3095)

IHC-P - High definition dendritic staining with ab3095 (very limited cell soma staining is seen).

Clonality Applications tested Host Species reactivity Datasheet M IHC-fr, IF Mm Hu, Mm, Rt, C. www.abcam.com/ab3095 Does not react with Ch.

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

NSE P WB Ch Hu, Mm, Rt ab14312 NSE P WB Rb Hu, Mm, Rt ab16873 NSE P IHC-Fr, IHC-P Rb Hu, B ab834 NSE [18H2] M ELISA, IP, WB Mm Hu, Mm. Does not react with Rt ab21253 NSE [37E4] M ELISA, WB Mm Hu, Mm, Rt ab16807 NSE [85F11] M ELISA, WB Mm Hu, Mm. Does not react with Rt ab16808 NSE [MIG-N3] M IHC-P, WB Mm Hu ab8197 NSE [V1-H14] M IHC-Fr, IHC-P Mm Hu ab790 PGP95 P IHC-Fr Gp Hu, Mm, Rt ab10410 PGP95 P IF, IHC-Fr, WB Rb Hu, Mm, Rt, Pi ab10404 PGP95 P WB Gt Hu ab11145 PGP95 P IHC-P Rb Hu ab15503 PGP95 [13C4 / I3C4] M IF, IHC-F, IHC-Fr, IHC-P Mm Hu, Rt, Gp ab8189 PGP95 [31A3] M ELISA, IHC-P, WB Mm Hu, Rt ab20559 Plexin A1 P ELISA, WB Rb Hu, Mm, Rt ab23391 SOX2 P IF Rb Hu, Mm ab15830 SOX22 P IHC, IP, WB Rb Hu, Mm, Rt ab12062 STEP / PTPN5 P WB Rb Hu, Mm ab12151 STEP / PTPN5 [23E5] M WB Mm Rt ab16967 STEP / PTPN5 peptide (phosphatase domain) BL ab22661 Neuronal markers Survivin P WB Rb Hu ab8228 Survivin P AF, H, ICC, IF, IHC, IHC-F, IHC-Fr, IHC-P, IP, WB Rb F, D, Hu, Mm ab469 Survivin [321] M IF, IP, WB Mm Hu ab10584 Survivin [6011] M IF, WB Mm Hu, Mm, Rt ab4470 Survivin [8E2] M IF, IHC-P Mm Hu, Rt ab3258 Survivin (phospho T34) P WB Rb Hu ab10720 Survivin peptide BL ab7865 Survivin peptide BL ab7866 Tyrosine Hydroxylase P ELISA, IHC-Fr Rb Rt ab6211 Tyrosine Hydroxylase P ICC, IP, WB Sh Ma ab113 Tyrosine Hydroxylase P ICC, IF, IHC, IHC-Fr, IP, WB Rb Ma ab112 Tyrosine Hydroxylase [185] M IHC-Fr, WB Mm Hu, Mm, Rt ab10372 Pep. avail. Tyrosine Hydroxylase [300-108] M ICC, IHC, IP, WB Mm Ma ab111 Tyrosine Hydroxylase [GD7] M IF, IHC-Fr, WB Mm Hu, Mm, Rt ab23763 Tyrosine Hydroxylase (phospho S19) P IF, IHC-Fr, WB Rb Hu, Mm, Rt ab23784 Tyrosine Hydroxylase (phospho S31) P IF, IHC-Fr, WB Rb Mm, Rt, Hu ab23782 Pep. avail. Tyrosine Hydroxylase (phospho S40) P IF, IHC, WB Rb Hu, Mm, Rt, Mk, Pi, Q ab16557

NeuN antibody [4G2] (ab13938)

IHC - ab13938 at a dilution of 1/1000, staining NeuN on rat brain tissue (30µm thick coronal sections). Images showing neuronal staining.

Clonality Applications tested Host Species reactivity Datasheet M WB, IHC-P, IHC-Fr, IF, ICC Mm Mm, Rt www.abcam.com/ab13938

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Neuroscience > Neuronal Markers > Neuron > Synapse

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

14-3-3 P IP, WB Rb Hu, Mm, Rt ab9063 14-3-3 P IHC-P, IP, WB Rb Hu, Mm, Rt ab6081 14-3-3 [2Q248] M WB Mm Rt ab14121 14-3-3 [3F7] M IP, WB Mm Hu ab14108 14-3-3 beta P IHC-P, IP, WB Rb Hu, Mm, Rt ab15260 14-3-3 beta P WB Rb Hu, Mm, Rt ab16730 14-3-3 beta [60C10] M ELISA, WB Mm Hu, Mm, Rt ab16859 14-3-3 beta, prediluted P IHC-P Rb Hu, Mm, Rt ab15262 14-3-3 beta + epsilon + zeta [3C8] M WB Mm Hu, Mm, Rt ab12346 14-3-3 beta + zeta P WB Rb Ma, Dm ab14113 14-3-3 beta + zeta [22-IID8B] M ICC, IHC, WB Mm Hu, Mm, Rt, B, D, Mk, Pi, Rb, Sh ab12347 14-3-3 eta + gamma + zeta P ELISA, WB Rb Many ab14124 14-3-3 gamma (acetyl V1) [KC21] M WB Mm Hu ab17005 14-3-3 gamma [3F8] M IP, WB Mm Hu ab14117 14-3-3 sigma [1N6] M IF, IHC-P, IP, WB Mm Hu ab14123 14-3-3 tau [3B9] M IP, WB Mm Hu, Mm, Rt, B ab10439 14-3-3 Binding Motif (phospho S) P ELISA, IHC-P, IP, WB Rb Many ab14127 14-3-3 Binding Motif (phospho S) [3i2] M ELISA, IP, WB Mm Many ab14126 Amphiphysin [8] M IHC, IP, WB Mm Hu, Mm, Rt, B ab12232 Amphiphysin [C14-23] M ELISA, ICC, IHC, IP, WB Mm Hu, Mm, Rt ab16770 Pep. avail. C3 P IHC-Fr, WBRb Mm ab11887 C3 P IF Sh Hu ab14396 Pep. avail. C3 P ELISA, WBCh Hu ab14232 C3 P DID Gt Hu ab13329 C3 P Ie Gt Hu ab20069 Pep. avail. C3 P IF Gt Gp ab20423 C3 P ELISA, ID, WB Rb Hu, Mm, Rt ab23891 C3 [004-01] M ELISA, WB Mm Hu ab17455

Neuronal markers C3 [118-02] M ELISA Mm Rt ab17456 C3 [11H9] M FACS, IHC-Fr, IP, WB Rt Mm ab11862 Calpastatin P ELISA, WBCh Hu ab16423 Calpastatin [2G11D6] M ICC, WB Mm Hu, Rt, C, Pi ab3515 CASK P IHC, WB Rb Hu, Rt, Mm ab3383 CASK P IP, WB Rb Mm, Rt, D ab11343 CD172a [BL1H7] M FACS Mm Pi ab23772 Cellubrevin P IF, WBRb Rt. Does not react with Hu or D ab2102 Cellubrevin P IF, WB Rb Hu, Mm Does not react with Rt or D ab2103 CPEB3 P Fast track Rb Fast track ab12024 CPEB3 P WB Rb Mm ab10883 CSP P IHC, IP, WB Rb Mm, Rt, X, C ab13250 Dynamin [0T178] M ICC, IP, WB Mm Hu, Mm, Rt, B ab14447 Dynamin [0T179] M ELISA, IHC-P, IP, WB Mm Hu, Mm, Rt, B ab14450 Dynamin [D5] M ICC, IF, IP, WB Mm Hu, Mm, Rt ab12429 Dynamin (phospho S774) P IF, IHC, WB Sh Hu, Mm, Rt ab18090 Dynamin (phospho S778) P WB Sh Hu, Rt ab18101 Dynamin 1 P WB Rb Rt ab3456 Dynamin 1 [4E67] M IF, IP, WB Mm B, Ch, Hu, Mm, Rt ab14448 Dynamin 1 [D5] M IHC, IP, WB Mm Hu, Mm, Rt, C, Ch ab13251 Dynamin 2 P WB Rb Hu, Rt, Mm ab3457 Dynamin 3 P WB Rb Hu, Rt, Mm ab3458 Homer (1a+1b+1c) P IHC-Fr, WB Rt Hu, Mm, Rt ab11157 Homer (1b+1c) P ELISA, FACS, ICC, IF, IHC-Fr, IP, RIA, WB Rt Hu, Mm, Rt ab18550 LAR P WB Rb Hu ab10557 LAR peptide BL ab22637 Mint1 P WB Rb Rt ab3448 MYRIP P Fast track Gt Fast track ab10149 PSD93 P IHC-Fr, WBRb Rt ab2930

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

PSD93 P IHC-P, WB Gt Mm ab12097 PSD95 P IHC-P, WB Gt Mm ab12093 PSD95 [6G6-1C9] M IF, IP, WB Mm Rt ab2723 PSD95 [7E3-1B8] M ICC, IP, WB Mm Mm, Rt, B ab13552 PSD95 [OT123] M IHC-Fr, IP, WB Mm Hu, Mm, Rt ab18359 Rab4 P ICC, IF, IP, WB Rb Hu, Mm, Rt, Ch, D, Fi, Gp, Ha, Mk, Pi, Rb, Sh ab13252 Rab5 P ICC, IHC, IP, WB Rb Hu, Mm, Rt, B, D, Gp, Ha, Mk ab13253 Rabphilin 3A P WB Rb Mm, Rt, B ab12234 rSec6 [9H5] M IHC, WB Mm Hu, Mm, Rt, B, Ch, D, Ha, Mk, Pi, Rb, Sh ab12235 rSec8 [14G1] M IP, WB Mm Hu, Mm, Rt, B, Ch, D, Gp, Ha, Mk, Pi, Rb, Sh ab13254 SAP102 P IHC-Fr, WB Rb Rt. Does not react with Mm ab3438 SAP102 P WB Gt Mm ab12086 Pep. avail. SIRP alpha P WB Rb Hu, Rt, Mm ab8120 SIRP alpha P IHC-P, WBRb Hu, Mk. Does not react with Mm ab2971 SIRP [MRC OX-41 ] M FACS, IHC-Fr, IHC-P, WB Mm Rt ab9295 SNAP23 P WB Rb Rt, Mmab3340 SNAP23 P IF, WB Rb Hu, Rt ab4114 Neuronal markers SNAP23a P WB Rb Hu, B, D, Ha ab13804 SNAP25 [4H251] M WB Mm Hu, Mm, Rt, X, B, Ha, In, Pi ab18002 SNAP25 [SP12] M ELISA, IHC-P, WB Mm Hu, Ha, Rt, Pi ab11102 SNAP91 [2D11] M IHC, IP, WB Mm Hu, Mm, Rt, B ab12237 SNAP91 [AP180-I] M AP, ICC, IP, WB Mm Hu, Rt, B ab11329 Synapsin I P AF, IF, IHC-F, IHC-Fr, IP, WB Rb B, Gp, Hu, Mm, Rt ab8 Synapsin I (phospho S603) P Dot, WB Rb Rt ab13879 Synapsin I (phospho S9) P WB Rb Hu, Mm, Rt, X , B ab16554 Synapsin II M IP, WB Rb Mm, Rt, B ab12240 Synapsin II P WB Rb Mm, Rt ab13258 Synaptobrevin [SP10] M WB Mm Hu, Rt, Ha, Pi ab11109

SAP97 antibody (ab3437)

IF - Free-floating staining of rat brain tissue with ab3437 (used at 1/3000 overnight).

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-Fr, IHC Rb Rt www.abcam.com/ab3437

SNAP25 antibody (ab5666)

IF - SNAP-25 immuno staining in cultured rat hippocampal cells using ab5666 (1/100) yields a pattern consistent with cytoplasmic and plasma membrane staining.

Clonality Applications tested Host Species reactivity Datasheet P WB, IF Rb Mm, Rt, C www.abcam.com/ab5666

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Synaptophysin [SP15] M ELISA, IHC-P, WB Mm Hu, Rt, Ha, Pi ab11105 Synaptophysin [SVP38] M FACS, ICC, WB Mm Rt ab12353 Synaptophysin [SY38] M IHC-Fr, IHC-P Mm Rt, Mm, B ab8049 Synaptobrevin [SP11] M ELISA, IHC-P, WB Mm Hu, Pi. Does not react with Rt ab11104 Synaptophysin P IHC-F, IHC-Fr, IHC-P Rb Hu ab7837 Synaptophysin P IHC-Fr, IP, WB Rb Hu, Mm, Rt ab14692 Synaptophysin [4E206] M IHC, IP, WB Mm Hu, Mm, Rt, B ab18008 Synaptophysin [SP11] M IHC-P, WB Rb Hu ab16659 Synaptotagmin P ELISA, WB Ch Hu, B, Mm, Pi, Rt ab8037 Synaptotagmin P IF, WB Rb Hu, Rt ab10104 Synaptotagmin P ICC, IP, WB Rb Rt, X ab13260 Synaptotagmin [ASV30] M ICC, IHC, IP, WB Mm Mm, Rt, X, B, Ch, Rb, Sha ab13259 SynCAM P WB Rb Rt ab3910 Syndecan P IHC-P Rb Hu ab16037 Syndecan [BC/B-B4] M IHC-Fr, IHC-P Mm Hu ab714 Syntaxin P ELISA, IP, WB Ch Hu, B, Mm, Pi, Rt ab8038 Pep. avail. Syntaxin P WB Rb Mm, Rt, C ab13262 Syntaxin [SP6] M IHC-Fr, IP, WB Mm Hu, Mm, Rt, X, B, F, Ha, Pi ab12236 Syntaxin [SP8] M ELISA, IHC-Fr, WB Mm Hu, Rt, Pi, Ha ab257 Syntaxin [STX01 (HPC-1)] M IHC-P, WB Mm Hu, Rb, Rt, C. Does not react with Gp ab3265 Syntaxin 2 P IP, WB Rb Hu, Mm, Rt, X, B, Dm, Ha, Mk, Pi, Rb, Sh ab12369 Syntaxin 3 P IF, WB Rb Ma ab4113 Syntaxin 13 [15G2] M ICC, WB Mm Mm, Rt, D, Ha ab13261 Pep. avail. VAMP1 P IP, WB Rb Hu, Rt, Mm, D ab3346 Pep. avail. VAMP2 P IP, WB Rb Rt, Mm ab3347

Neuronal markers VAMP2 P ICC Ch Hu, Mm, Rt ab14279 VAMP8 P IF, IM, IP, WB Rb Hu, Rt, D ab6186

View the synapse. Neuroscience Antibody Locator at www.abcam.com/ neuroscience

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Neuroscience > Neurotransmission > Calcium Signalling > Calcium Binding Proteins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Calbindin P ELISA, IF, IHC-P, IP, Radio-IHC, WB Rb Hu, Rt ab11426 Calbindin [CL-300] M IF, IHC-Fr, IHC-P Mm Hu, Ch, Fi, Gp, Ha, Mk, Mm, Rb, Rt, Sh ab9481 Calretinin P IP, WB Rb Hu, Mm, Rt ab14689 Pep. avail. Calretinin P IHC-Fr, IHC-P Rb Hu, F, Ch, Mm, Rt, Re ab702 Calretinin [CRT01] M IHC-Fr, IHC-P Mm Mm ab3934 Frequenin P Dot, ELISA, IHC-F, IM, WB Ch Most Ma ab7623 NCS1 P WB Rb Rt ab5807Pep. avail. Parvalbumin P ELISA, IHC-P, IP, WB Rb Hu, Rt ab11427 Pep. avail. S100 P IF, IHC-P Rb Hu, Rt, Mm, C, Pi ab868 S100 P IF, IHC-F, IHC-Fr, IHC-P Rb Hu, Rt, B ab7853 S100 [4E3 ] M IA, P, WB Mm Hu ab10202 S100 [6G1] M ELISA, IA, P, WB Mm Hu ab10203 S100 [8B10] M EIA, ELISA, IF, IHC-P, WB Mm Hu, Rt ab8330 S100 [B321] M IF, IHC, WB Mm Hu, Rt, Mm, C, Pi ab7852 Pep. avail. S100A8 + S100A9 [27E10] M FACS, IA, IHC-Fr, IHC-P, IP, WB Mm Hu, Mk ab17050

NCS1 antibody (ab18060)

IF - a cross section of an E14 rat spinal cord at the cervical enlargement labeled with chicken anti-NCS-1 (ab18060; 1/100). Neurotransmission

Clonality Applications tested Host Species reactivity Datasheet P Conjugation, ELISA, IHC, WB Ch Hu, Mm, Rt www.abcam.com/ab18060

Neuroscience > Neurotransmission > Calcium Signaling > Calcium Channels > L-type

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Calcium channel L type DHPR alpha 2 subunit [20A] M IHC-Fr, WB Mm Hu, Rb, Rt, Mm, Gp ab2864 DHPR alpha 1 P IP, WB Sh Rb ab14446 DHPR alpha 1 [1A] M IHC-Fr, IP, WB Mm Hu, Rb, Rt, Mm, Gp ab2862 pan Calcium Channel P ICC, IP, WB Rb Mm, Rt ab6298

Neuroscience > Neurotransmission > Calcium Signaling > Calcium Channels > N-type

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

CACNB3 P IF, IP, WB Rb Rt ab16717 pan Calcium Channel P ICC, IP, WB Rb Mm, Rt ab6298 Pep. avail.

Neuroscience > Neurotransmission > Calcium Signaling > Calcium Channels > P / Q / R -type

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/... pan Calcium Channel P ICC, IP, WB Rb Mm, Rt ab6298

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Neuroscience > Neurotransmission > Calcium Signaling > Calcium Channels > Ryanodine Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Ryanodine Receptor [34C] M IHC-Fr, IHC-P, IP, WB Mm Hu, Rb, Rt, Mm, B, D, Sh, Fi, Am ab2868 Pep. avail. Ryanodine Receptor [C3-33] M FACS, IHC-P, IP, WB Mm Rb, Rt, D, Gp, Ch, Fi, Am ab2827

Neuroscience > Neurotransmission > Calcium Signaling > Calmodulin / CaMK

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

CABP P WB Gt Hu ab2936 CaMKI P IP, WB Rb Hu, Mm, Rt ab15810 CaMKII P WB Rb Hu, Mm, Rt, Rb, B, Ch, Gp, Ha ab19223 CaMKII [6G9] - Azide free M IHC, RIA, WB Mm Mm, Rt, B ab22609 CaMKII (phospho T305) P WB Rb Rt ab22183 CaMKII alpha P WB Rb Mm, Rt ab3908 CaMKII alpha (phospho T286) P WB Rb Mm ab5683 CaMKII alpha (phospho T286) P WB Rb Hu ab18317 CaMKII alpha (phospho T286) [22B1] M ELISA, IF, IP, WB Mm Rt ab2724 CAMKIV P IHC-Fr, WB Rb Hu, Rt, Mm, D, Mk ab3557 CAMKIV P IP, WB Rb Rt ab22658 CaMKK P WB Rb Hu, Mm, Rt ab22655 Neurogranin P ELISA, ICC, IHC, IP, WB Rt Rt, Mm ab23570

CaMKII alpha antibody [6G9] (ab2725)

IF - Cytoplasmic staining with ab2725 in rat dorsal horn of the spinal cord (lamina II).

Neurotransmission Clonality Applications tested Host Species reactivity Datasheet M IF, IP, IHC, WB Mm Rt www.abcam.com/ab2725

Neuroscience > Neurotransmission > Calcium Signaling > IP3R

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

IP3R1 P IHC, IP, WB Rb Rt, D ab5908

Neuroscience > Neurotransmission > Intracellular Signaling > Adapters

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

AP3B2 P ICC, IHC Ch Hu, Mm, Rt ab14227 CAPON P Fast track Gt Fast track ab3829 GABARAP P WB Rb Hu ab1133 PATJ P Fast track Gt Fast track ab8225 PDZK3 P Fast track Gt Fast track ab6037 PSD P Fast track Gt Fast track ab5962 PSD93 P IHC-Fr, WBRb Rt ab2930 PSD93 P IHC-P, WBGt Mm ab12097 PSD95 P IHC-P, WBGt Mm ab12093 PSD95 [6G6-1C9] M IF, IP, WB Mm Rt ab2723

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

PSD95 [7E3-1B8] M ICC, IP, WB Mm Mm, Rt, B ab13552 PSD95 [OT123] M IHC-Fr, IP, WB Mm Hu, Mm, Rt ab18359 SAP102 P WB Gt Mm ab12086 SAP102 P IHC-Fr, WBRb Rt. Does not react with Mm ab3438 SAP97 P IHC-Fr, WBRb Rt ab3437 SHANK1 P Fast track Gt Fast track ab10006 SHANK1 P ELISA Rb Rt ab18001 SHANK1 P WB Rb Hu ab3896 SHANK1a P IF, WBRb Hu ab3897 SNTG1 P Fast track Gt Fast track ab10079 SynGAP P ICC, WB Rb Rt, Mm ab3344 SynGAP P ICC, IP, WB Rb Rt ab17914

Neuroscience > Neurotransmission > Intracellular Signaling > Cytoskeletal

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

CR16 P WB Gt Mm ab2238 Neurotransmission Neurabin 1 P IF, WB Rb Hu, Rt ab3483 Neurabin 2 P IP, WB Rb Hu, Mm, Rt, Mk, D ab18561 Neurofibromin P IP, WBRb Hu ab17963 Neurofibromin [McNFn27] M AP, ELISA, IHC-P, IP, WB Mm Hu, Mm, Rt ab8130 Neurofibromin [PcNFn27] P IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt ab8132 SNTG1 P Fast track Gt Fast track ab10079

Neuroscience > Neurotransmission > Intracellular Signaling > Kinases

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

ALK c [ALKc] M IHC-Fr, IHC-P Mm Hu ab650 Casein Kinase 1 delta P ELISA, IHC-P Gt Hu ab10877 CASK P IHC, WB Rb Hu, Rt, Mm ab3383 CASK P IP, WB Rb Mm, Rt, D ab11343 Pep. avail. COPS3 P WB Rb Hu ab10463 COPS4 [BL1061] P WB Rb Hu ab12322 Pep. avail. CrkL P WB Gt Hu, Mm ab10907 CSN1 P WB Rb Hu ab10413 CSN2 P WB Rb Hu ab10426 CSN5 [BL1062] P WB Rb Hu ab12323 CSN7b P WB Rb Hu ab11895 EphA1 P ELISA, WBRb Hu ab5385 EphA1 P I-ELISA, WBGt Hu ab7036 EphA2 P ELISA, WB Rb Ha, Hu, Mm ab5387 EphA3 P I-ELISA, WB Gt Mm ab7038 EphA3 [III-A4] M FACS, IP Mm Hu ab19439 EphA4 P ELISA, WB Rb Hu ab5389 EphA4 P I-ELISA, WBGt Mm ab7039 EphA5 P ELISA, WB Rb Hu, Mm ab5397 EphA5 P ELISA, WB Gt Mm, Rt ab10612 EphA6 P ELISA, WBGt Mm ab10613 EphA7 P ELISA, WBRb Hu ab5411 EphA7 P ELISA, WB Gt Mm ab10614 EphA8 P ELISA, WB Gt Mm ab10615 EphB1 P ELISA, WB Rb Hu, Mm ab5414 EphB1 P I-ELISA, WB Gt Rt ab7042

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

EphB1 P Dot, IP Sh Hu ab10406 EphB2 P ELISA, WB Rb Hu, Mm ab5418 EphB2 P Dot, ELISA Gt Mm ab10616 EphB3 P ELISA, WBGt Mm ab10617 EphB4 P Dot, ELISA, Inhib Gt Mm ab10618 EphB6 P Dot, ELISAGt Mm ab10619 Ephrin A2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610 Ephrin A3 P ELISA, WB Gt Hu. Does not react with Mm ab10611 Ephrin A4 P I-ELISA, WB Gt Hu ab7041 Ephrin B3 P ELISA, WB Gt Hu ab7044 HUNK P ICC, IF, IHC, IHC-P Rb Hu, Mm ab13117 Neurogranin P ELISA, ICC, IHC, IP, WB Rt Rt, Mm ab23570

Neuroscience > Neurotransmission > Intracellular Signaling > Phosphatases

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

PP2A alpha and beta P WB Gt Hu ab1309 PP2A alpha and beta P ICC, WB Sh Hu, Mm, Rt, B ab16450 PPP2R5A P ICC, WB Rb Hu, Mm, Rt, B ab18136 PPP2R5A P WB Gt Hu ab1084 PPP2R5A P WB Sh Hu, Mm, Rt ab16449 PPP2R5B P WB Gt Mm ab1366 PPP2R5C P ICC, WB Rb Hu, Mm, Rt, B ab18133 PPP2R5D P WB Gt Hu ab1361 PPP2R5E P WB Gt Mm ab1083 Neurotransmission

Neuroscience > Neurotransmission > Intracellular Signaling > Regulation

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

CSN1 P WB Rb Hu ab10413Pep. avail. CSN2 P WB Rb Hu ab10426 CSN5 [BL1062] P WB Rb Hu ab12323 CSN7b P WB Rb Hu ab11895 p35 P WB Rb Hu ab10570 p35 [ES19] M IP Mm Hu ab3222 p35 nck5a [SPM430] M IP Mm Hu ab19933 Phosphoserine P ELISA, IP, WB Rb Many ab9332 Phosphoserine (HRP) P ELISA, IP, WB Rb Many ab9334 Phosphoserine [3C171] M ELISA, WB Mm Many ab17465 Phosphoserine [4A9] M ELISA, IP, WB Mm Many ab21642 Phosphoserine [PSR-45] M I-ELISA, WB Mm Many ab6639

Phosphoserine/threonine/tyrosine antibody [SPM101] (ab15556)

IHC - Human breast carcinoma stained with ab15556 (FFPE-sections).

Clonality Applications tested Host Species reactivity Datasheet M WB, IP, ELISA, IHC-P, IF Mm Many www.abcam.com/ab15556

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 46 Neuroscience resources: www.abcam.com/neuroscience Neuroscience > Neurotransmission > Intracellular Signalling > Regulation

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Phosphoserine/threonine P ELISA, IP, WB Rb Many ab17464 Phosphothreonine-Proline / Phosphoserine-Proline P ELISA, WB Rb Many ab9344 Phosphothreonine-Proline / Phosphoserine-Proline (HRP) P ELISA, WB Rb Many ab9345 Phosphotyrosine P ELISA, WBRb Many ab17301 Phosphotyrosine P Dot, ELISA, IF, WB Rb Many ab17303 Phosphotyrosine P ELISA, IP, WB Rb Many ab9329 Phosphotyrosine P ELISA, IP, WB Rb Many ab9319 Phosphotyrosine (AP) M ELISA, RIA Mm Many ab2339 Phosphotyrosine [1BB49] (Biotin) M IHC, WB Mm Many ab17371 Phosphotyrosine [1BB49] (FITC) M IHC, WB Mm Many ab17372 Phosphotyrosine [2Q258] (HRP) M ELISA, WB Mm Many ab17288 Phosphotyrosine [P9V6] (Biotin) M IA, WB Mm Many ab7227 Phosphotyrosine [PY20] (Biotin) M ELISA, ICC, WB Mm Many ab16388 Phosphotyrosine [IG2] M FACS, ICC, IP, WB Mm Many ab5592 Phosphotyrosine [P9V6] M IA, WB Mm Many ab7226

Phosphotyrosine antibody [PY20] (ab10321)

IF - in the rat cortex. Shows microglial cells stained. IHC free-floating protocol using 4% PFA fixed brain tissue. Neurotransmission Primary antibody ab10321 was used at 1 ug/ml incubated overnight at room temperature.

Clonality Applications tested Host Species reactivity Datasheet M AP, ELISA, FACS, IF, IHC-FrFl, IP, WB Mm Rt www.abcam.com/ab10321

Neuroscience > Neurotransmission > Neurotransmitters > Acetylcholine

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Acetylcholine P ELISA, ICC, IHC-P Rb Many ab8884 Acetylcholine M ICC, IHC-P Mm Many ab6470 Acetylcholinesterase P ELISA, IP, Neut, PCC, WB Gt B ab7396 Acetylcholinesterase (HRP) P ELISA, IHC-Fr, IM, PCC, WB Gt B ab7397 Acetylcholinesterase [190-01] M ELISA, WB Mm Hu, B ab17774 Acetylcholinesterase [HR2] M ELISA, IHC-Fr, IP Mm Hu, Rb, B, F, Gp ab2803 Acetylcholinesterase [HYB 111-05] M ELISA, WB Mm Hu. Does not react with Fi ab23455 Acetylcholinesterase [ZR3] M IHC-Fr, IP Mm Rb, Rt, F, Gp ab2802 Choline Acetyltransferase P IHC-F, WB Sh Hu, Rt, Rb, Gp, Pi ab18736 Choline Acetyltransferase P ELISA, IHC Rb Mm, Pi, Rt ab6168 PBP P WB Gt Hu ab2634

Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Aspartate

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Glutamate Aspartate Transporter P FACS, ICC, IHC, PCC, WB Rb Rt ab416 L-Aspartate P ICC, IHC-F, IHC-Fr, IM Rb Many ab9439

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Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > GABA

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

GABA A Receptor alpha 1 P IP, WB Rb Rt ab5781 GABA A Receptor alpha 2 P WB Rb Mm, Rt ab8342 GABA A Receptor alpha 4 P IP, WB Rb Mm, Rt ab4120 GABA A Receptor alpha 4 P Dot, WB Rb Rt ab13872 GABA A Receptor alpha 5 P WB Rb Rt, Mm ab3700 GABA A Receptor alpha 6 P IP, WB Rb Mm, Rt ab5365 GABA A Receptor beta 1 P IP, WB Rb Mm, Rt ab16703 GABA A Receptor beta 1 P WB Rb Hu ab23338 GABA A Receptor beta 2 P IP, WB Rb Mm, Rt ab8340 GABA A Receptor beta 3 P IP, WB Rb Mm, Rt ab4046 GABA A Receptor delta P WB Rb Hu, Mm ab10100 GABA A Receptor gamma 2 P IP, WB Rb Mm, Rt ab4073 GABA P ICC, IHC-F, IHC-Fr, IM Rb Many ab9446 GABA P IF, IHC Gp Many ab17413 GABA P ELISA, IF, IHC Rb Rt ab17390 GABA [2Q67] M IHC-Fr, IHC-P Mm Hu ab17313 GABA B Receptor 1 P WB Rb Hu ab1135 GABA B Receptor 2 (phospho S892) P IF, IHC, WB Rb Mm, Rt ab3900 GABA B Receptor 2 P WB Rb Hu ab1134 GABA Transporter 1 / GAT 1 P IF, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch ab426 GABA Transporter 2 P IF, IHC-Fr, WB Rb Hu, Mm, Rt, Ch ab2896 GABA Transporter 3 / GAT 3 P IF, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch ab431

Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Glutamate

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/... Neurotransmission GAD65 P IHC-Fr, IHC-P Rb Hu, Rt ab2348 GAD65, prediluted P IHC-P Rb Hu, Rt ab15299 Glutamate P ICC, IHC-F, IHC-Fr, IM Rb Many ab9440 Glutamate P ELISA, ICC Rb Many ab8889 Glutamate M ELISA, IHC-P Mm Many ab8893 Glutamate [2Q71] M IHC Mm Rt ab17463 Glutamate Aspartate Transporter P FACS, ICC, IHC, PCC, WB Rb Rt ab416 Glutamate Transporter 1 P FACS, ICC, IF, IHC, PCC, WB Rb Hu, Rt ab417 Glutamate Receptor 6+7 (metabotropic) P IHC-P, IP, WB Rb Hu, Mm, Rt, Ch, Mk ab15307

Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Glycine

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Glycine P ICC, IHC-F, IHC-Fr, IM Rb Many ab9442 Glycine Receptor alpha 1 P IP, WB Rb Many ab475

Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Related targets

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

5-hydroxy Tryptamine P ELISA, IHC-Fr Rb Many ab6458 6-hydroxy Tryptamine P ELISA, IHC-P Rb Many ab6455 ARMET P WB Gt Hu ab1347 ARMET peptide (222-234) ab22811 Taurine P ELISA, IHC-FrRb Many ab6467 Taurine P ICC, IHC-F, IHC-Fr, IM Rb Many ab9448 Tyramine P ELISA Rb Many ab6451

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Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Tryptophan

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

5-hydroxy Tryptophan P ELISA, ICC, IHC-Fr Rb Many ab8890 5-hydroxy Tryptophan [4D18] M ELISA Mm Many ab18171 5-methoxy Tryptophan P ELISA Rb Many ab6476 Tryptophan P ELISA, ICCRb Many ab8886 Tryptophan Hydroxylase P IF, IHC, WB Sh Hu, Rt ab3907 Tryptophan Hydroxylase (phospho S58) P WB Rb Hu, Mm, Rt, X ab23779

Neuroscience > Neurotransmission > Neurotransmitters > Biogenic Amines > Dopamine

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

6-hydroxy Dopamine P ELISA Rb Many ab9429 DOPA Decarboxylase P ICC, WB Rb Rt ab3905 Dopamine M BL, IHC-Fr, IHC-P Mm Many ab1001 Dopamine P ELISA, IHC Rb Many ab6427 Dopamine P ELISA, ICC Rb Many ab8888 Pep. avail. Dopamine M ELISA, ICC, IHC-Fr, IHC-P Mm Many ab8892 Pep. avail. Dopamine [2B11] M ICC, Neut Mm Many ab8281 Pep. avail.

Dopamine beta Hydroxylase P IHC, WB Sh B, Hu, Rb, Mk. Neurotransmission Does not react with D, Rt or Sh ab6487 Pep. avail. Dopamine beta Hydroxylase P IF, IHC-P, WB Sh Hu, Mm, Rt, B ab19353 Dopamine Transporter P ELISA, WB Rb Hu, Mm, Rt ab18548 Dopamine Transporter [hDAT-LOOP] M ICC, IHC-Fr, WB Rt Hu, Rt ab5981 L Dopa M ELISA Mm Many ab5979 L Dopa P ELISA, IHC-Fr Rb Many ab6426 Methamphetamine [4E2] M ELISA Mm Many ab23328 Neurotensin P IHC-Fr, IHC-P, IHC-R Rb Ma ab11142 Tyrosine Hydroxylase (phospho S19) P IF, IHC-Fr, WB Rb Hu, Mm, Rt ab23784 Tyrosine Hydroxylase (phospho S31) P IF, IHC-Fr, WB Rb Mm, Rt, Hu ab23782 Tyrosine Hydroxylase (phospho S40) P IF, IHC, WB Rb Hu, Mm, Rt, Mk, Pi, Q ab16557 Tyrosine Hydroxylase P ICC, IF, IHC, IHC-Fr, IP, WB Rb Many ab112 Tyrosine Hydroxylase P ICC, IP, WB Sh Many ab113 Tyrosine Hydroxylase P ELISA, IHC-Fr Rb Rt ab6211 Tyrosine Hydroxylase [185] M IHC-Fr, WB Mm Hu, Mm, Rt ab10372 Tyrosine Hydroxylase [300-108] M ICC, IHC, IP, WB Mm Ma ab111 Tyrosine Hydroxylase [GD7] M IF, IHC-Fr, WB Mm Hu, Mm, Rt ab23763

Neuroscience > Neurotransmission > Neurotransmitters > Biogenic Amines > Noradrenaline

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Noradrenaline P ELISA, IHC-Fr Rb Many ab6454 Noradrenaline P ELISA, ICC, IHC-Fr Rb Many ab8887

Neuroscience > Neurotransmission > Neurotransmitters > Biogenic Amines > Related targets Amines

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

5-methoxy Tryptamine P ELISA, IHC-Fr Rb Many ab6458 6-hydroxy Tryptamine P ELISA, IHC-P Rb Many ab6455 CSPS P WB Rb Hu, Mm, Rt ab16870 CSPS [3F10] M ELISA, IP Mm Hu, Mm, Rt ab16773 Tyramine P ELISA Rb Many ab6451

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Neuroscience > Neurotransmission > Neurotransmitters > Biogenic Amines > Serotoin / 5HT

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

5 Methoxytryptamine P IHC Rb Rt ab15165 5HT1B Receptor P WB Rb Hu, Mm, Rt ab13896 Nitroserotonin P ELISA Rb Many ab6459 Serotonin P IHC-Fr, IHC-P Rb Hu, Rt ab10385 Serotonin P ELISA, ICC, IHC-Fr Rb Many ab8882 Serotonin [5HT-H209] M ICC, IHC-Fr, IHC-P Mm Hu, Mm, Rt, Gp ab16007 Serotonin [YC5/45] M IHC-P Rt Hu, Mm, Rt, Gp, Rb, X ab6336 Serotonin N-acetyltransferase P WB Rb Rt ab3505 Serotonin N-acetyltransferase (phospho S206) P WB Rb Rt ab3440 Serotonin N-acetyltransferase (phospho T29) P WB Rb Rt, Mm ab3439

Neuroscience > Neurotransmission > Neuropeptides > Hormones

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

ACTH P ICC, WB Ch Hu, Mm, Rt ab14064 ACTH P Dot, IHC-Fr Rb C, Rt ab8906 ACTH P IHC-P Rb Hu ab10335 ACTH P RIA Rb Hu, Rt, Mk ab11116 ACTH P IHC-Fr, IHC-P Rb Hu ab2407 ACTH [56] M ELISA, ICC, IHC-P Mm Hu, Rt ab21003 ACTH [A2H8] M ELISA, RIA Mm Hu ab11114 ACTH [A-4D5] M RIA Mm Hu ab11113 ACTH [B427] M IF, IHC-F, IHC-Fr, IHC-P, ICC, WB Mm Hu ab8615 BNP P ELISA, IHC, RIA Rb Hu, Rt. Does not react with Mm ab19645 BNP P ELISA, IF, RIA Rb Hu ab19646 Neurotransmission BNP [1B6] M EIA, ELISA, WB Mm Hu ab14699 BNP [2d3] (HRP) M EIA, ELISA Mm Hu ab14701 CGRP P IHC-F Sh Hu, Rt ab22560 CGRP P RIA Gt Hu ab14180 CGRP P IHC-Fr, IHC-P Rb Hu, Rt ab8056 CGRP [4901] M ELISA, ICC, Neut, RIA Mm Hu, Rt, D ab10987 Corticoliberin [4H9] M ELISA Mm Hu ab10034 Corticotropin Releasing Factor P AP, ICC, RIA Rb Hu, Rt ab8901 Estradiol P RIA Sh Many ab22441 Estradiol [6E1] M RIA Mm Hu ab20257 Estradiol [B42] M ELISA, IHC-Fr Mm Ma ab8743 Exendin 4 [ABS 012-20] M ELISA Mm Re ab23407 MC1 Receptor P IHC-P Rb Hu ab13033 MCH P ELISA, IHC-Fr Ch Hu, Mm, Rt ab15677 MSH gamma 1 P Dot, RIA, WB Rb C, Rt ab8910 Neurokinin B P IHC-P Rb Hu, Rt ab5772 Neuropeptide EI NH2 P IHC-Fr Rb Rt ab11141 Neuropeptide Y P IHC-Fr Gp Rt ab10341 Neuropeptide Y P Dot, IHC-Fr, RIA Rb Hu, Mm, Rt, F, B, Mk, Pi, Rb ab10980 Neuropeptide Y P IHC, IHC-F, IHC-Fr, IHC-P Sh Hu, Gp, Rb ab6173 Orexin A P ELISA, ICC, IHC-Fr Rb Hu ab16787 Orexin A P ELISA, RIA Rb Hu, Mm, Rt ab10981 Oxytocin P IHC-Fr Rb Rt ab11143 Oxytocin P IHC-P, RIA Rb Hu, Rt, Mm, Rb, Sh ab2078 Oxytocin Receptor P IHC-P Rb Hu ab13051 Peptide YY P IHC-Fr, IHC-P, IHC-R Rb Pi ab22663 Peptide YY P ELISA, WB Ch Hu, Mm, Rt ab15879 proBNP P RIA Gt Hu ab14352 proBNP [10h4] M EIA, ELISA, WB Mm Hu ab14703 proBNP [24E11] (HRP) M ELISA, WB Mm Hu ab13123 Somatostatin P IHC-F, IHC-Fr, IHC-P Rb Hu ab7840

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 50 Neuroscience resources: www.abcam.com/neuroscience Neuroscience > Neurotransmission > Neuropeptides > Hormones

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Somatostatin 14 P Dot, WB Rb Rt ab8903 Somatostatin 14, prediluted P IHC-P Rb Hu ab15522 Somatostatin 28 P EM, IF, IHC-Fr, IHC-P Rb Many ab22682 Somatostatin 28 P Dot, RIA, WB Rb Rt ab8904 Somatostatin Receptor P IHC-Fr, IHC-P Rb Rt, Mm ab2366 Somatostatin Receptor 1 P FACS, WB Rb Hu, Rt ab5827 Somatostatin Receptor 1 P IP, WB Rb Hu, Mm, Rt ab479 Somatostatin Receptor 2 P IF, IHC-P Rb Hu, Rt ab13120 Somatostatin Receptor 2 P IF, WB Rb Hu, Mm, Rt ab9550 Substance P P IHC-Fr Gp Hu, Mm, Rt ab10353 Substance P P RIA Rb Hu ab7305 Substance P P ELISA, WB Rb Many ab1123 Substance P [M09205] M IHC-Fr Rt Hu ab7340 Substance P [NC1/34 HL] M ELISA, H, ICC, IF, IHC, IHC-F, IHC-Fr, IHC-P, RIA Rt Av ab6338 Substance P [SP-DE4-21] M ELISA, IHC-P Mm Hu, Rt, Gp ab14184 Vasopressin P IHC-F, IHC-Fr, IHC-P Rb ab8745 VIP P IHC-Fr, IHC-P Rb Hu, Rt, Pi ab22736 VIP P IHC, IHC-F, IHC-Fr, IHC-P Sh Hu, Mm, Rt, B, D, Gp, Pi, Rb ab6184

Proteins Neurotransmission ACTH protein ab9586 Neuropeptide Y protein ab9599 Oxytocin protein ab9600 proBNP protein ab13125 Somatostatin 28 protein ab9606 Somatostatin protein ab9607 Substance P protein ab9608 VIP protein ab9609

Neuroscience > Neurotransmission > Neuropeptides > More Neuropeptides

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

FMRFamide P IHC-Fr Rb Rt ab10352 Neuropeptide EI NH2 P IHC-Fr Rb Rt ab11141 Neuropeptide FF1 P WB Rb Hu ab3898 Neuropeptide Y P IHC, IHC-F, IHC-Fr, IHC-P Sh Hu, Gp, Rb ab6173 Neuropeptide Y P IHC-Fr Gp Rt P ab10341 Neuropeptide Y P Dot, IHC-Fr, RIA Rb Hu, Mm, Rt, F, B, Mk, Pi, Rb ab10980 Neuropeptide Y CPON P IHC-P, IHC-R Rb Many ab22695 NPFF1 Receptor P ELISA, WB Rb Hu ab1401 NPFF1 Receptor P IHC-P Rb Hu ab13367 NPFF2 P WB Rb Hu ab3899 NPFF2 Receptor P ELISA, WB Rb Hu ab1402 NPFF2 Receptor P IHC-P Rb Hu ab13198 NPY5R P ICC, IHC-P Rb Hu ab13199 Orexin B P ELISA, RIA Rb Hu, Mm, Rt ab10982 Orexin B P ELISA, IHC, WB Rb Mm ab14970 PBP P WB Gt Hu ab2634 VRL1 P Dot, IHC-Fr, WB Rb Rt ab6183

Proteins Neuropeptide Y protein ab9599

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Neuroscience > Neurotransmission > Neuropeptides > Opioids

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Endomorphin 1 & 2 P ELISA, IHC-Fr Rb Hu, Mm, Rt, Mk, Pi ab10290 Endomorphin 1 P ELISA, IHC-Fr Rb Hu, Mm, Rt, Mk, Pi ab10285 Morphine M ELISA Mm Not applicable ab1060 Morphine [3A6] M ELISA Mm Not applicable ab23357 Nociceptin P IHC-Fr Gp Rt, Mk ab10276 Nociceptin P IHC-Fr Rb Rt, Mkab10277 Nociceptin P ELISA, I-ELISA, IHC-Fr, WB Rb Gp, Rt ab6174 NPFF2 P WB Rb Hu ab3899 Peptide E P IHC-Fr Rb Rt, B ab11144 ProDynorphin P IHC-Fr Gp Rt ab10280 ProDynorphin P IHC-Fr Rb Mm, Rt ab11137

Neuroscience > Neurotransmission > Neuropeptides > Tachykinins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Neurokinin 1 Receptor NK1 P FACS, WB Rb Hu ab466 Neurokinin 1 Receptor NK1 P AF, IHC-F, IHC-Fr Rb Gp, Mm, Rt ab6 Neurokinin 1 Receptor NK1 P IHC-Fr, WB Rb Rt, Mm, Gp ab8873 Neurokinin 1 Receptor NK1 P IF, IHC-P Rb Hu, Rt. Does not react with Gp ab13133 Neurokinin 3 Receptor NK3 P AF, IHC, IHC-F, IHC-Fr Rb Gp, Mm, Rt ab7 Neurokinin 3 Receptor NK3 P IHC-P Rb Hu ab13135 Neurokinin A Receptor P ICC, IHC-P Rb Hu ab13397 Neurokinin B P IHC-P Rb Hu, Rt ab5772 Substance P P ELISA, WB Rb Many ab1123 Pep. avail. Substance P P RIA Rb Hu ab7305 Neurotransmission Substance P P IHC-Fr Gp Hu, Mm, Rt ab10353 Substance P [M09205] M IHC-Fr Rt Hu ab7340 Substance P [NC1/34 HL] M ELISA, H, ICC, IF, IHC, IHC-F, IHC-Fr, IHC-P, RIA Rt Av, Ma ab6338 Substance P [SP-DE4-21] M ELISA, IHC-P Mm Hu, Rt, Gp ab14184 Thy12 [FF-10] M FACS, IHC-Fr, IP, WB Rt Mm ab22489

Proteins Substance P protein ab9608

Neuroscience > Neurotransmission > Nitric Oxide > Nitrated Species

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Nitrotyrosine P ELISA, IA, WB Gt Many ab20117 Nitrotyrosine P WB Rb Many ab23704 Nitrotyrosine P ELISA Ch Many ab2494 Nitrotyrosine [HM11] M ELISA, IHC-Fr, IHC-P, WB Mm Hu ab7048 NO Tyrosine P ELISA Rt Many ab6479 NO Tyrosine P ELISA Rb Many ab3916 NO2 Tyrosine P ELISA Rb Many ab6471 NO2 Tyrosine P ELISA Rt Many ab3805

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Neuroscience > Neurotransmission > Nitric Oxide > NOS

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/... bNOS P IHC-Fr, WB Rb Rb, Rt, Mm, B ab3511 bNOS P IF, IHC-F, IHC-Fr, IHC-P, WB Sh Gp, Rb, Rt ab6175 bNOS P WB Rb Hu, Rt ab19892 CAPON P Fast track Gt Fast track ab3829 DDAH2 P WB Gt Mm ab1383 Pep. avail. eNOS P ELISA, IP, WB Rb B ab3868 eNOS P ICC, WB Rb Hu, Mm, Rt, B ab4225 eNOS P IHC-P, WB Rb B, D, Hu, Mm, Pi, Rt ab5589 eNOS (phospho S116) P ELISA, IP, WB Gt B ab3867 iNOS P IHC-P, WB Rb Hu, Mm, Rt ab15323 iNOS P IF, WB Rb Hu, Rt, Mm ab3523 iNOS P ELISA, WB Rb Hu, Mm, Rt ab4226 NOS P IHC-P, WB Rb Hu, Mm, Rt, C, Pi ab15203 nNOS (neuronal) P WB Rb Hu ab3895 nNOS (neuronal) P WB Rb Rt ab5586 nNOS (neuronal) (phospho S1416) P WB Rb Rt ab5583 pan NOS P IHC-Fr, WB Rb Hu, B, Dm, Mm, Pi, Rt, Fi ab3342 pan NOS [NOS-3F7-B11 B5] M IHC-Fr, WB Mm Rt, Mm, B ab2801

nNOS antibody (ab1376) Neurotransmission

IHC-FrFl - ab1376 immunostaining nNOS in rat dorsal root ganglion neurons 2 weeks post spinal nerve ligation.The staining is restricted to the cytoplasm of a subpopulation of neurons.

Clonality Applications tested Host Species reactivity Datasheet P WB, IHC-FrFl Gt Mm www.abcam.com/ab1376

Neuroscience > Neurotransmission > Pharmacological Tools

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Amitriptyline [1BB831] M ELISA, RIA Mm Not applicable ab15176 Amitriptyline [202] - BSA and Azide free M IA Mm Not applicable ab20408 Amphetamine P ELISA, IF, IHC, IP, WB Gt Not applicable ab15208 Amphetamine [2DD1] M ELISA Mm Not applicable ab15210 Atrazine P ELISA Sh Not applicableab22435 Benzodiazepine P ELISA Sh Not applicableab15212 Ketanserin M ELISA Mm Not applicableab1061 Phenobarbital M ELISA Mm Not applicableab1097 Phenobarbital [F1#2G7A7] M EIA, RIA Mm Not applicable ab20871

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Acetylcholine

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Muscarinic Acetylcholine Receptor 1 P WB Rb Mm ab3480 Muscarinic Acetylcholine Receptor 2 P IHC-P Rb Hu ab13043 Muscarinic Acetylcholine Receptor 2 [31-1D1] M IP, WB Mm Hu, Rt, Mm, Pi. Does not react with Ch ab2805 Muscarinic Acetylcholine Receptor M3 P IHC-P Rb Hu ab13063 Muscarinic Acetylcholine Receptor M4 P IHC-P Rb Hu ab13065

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Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Adrenergenic Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

alpha 1b Adrenergic Receptor P WB Ch Hu ab15851 alpha 1b Adrenergic Receptor P IHC-P Rb Hu ab12933 alpha 1c Adrenergic Receptor P IHC-P Rb Hu ab12935 alpha 1d Adrenergic Receptor P IHC-P Rb Hu ab12923

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Cannabinoid Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Cable pilus P WB Rb Bu ab13961 Cannabinoid Receptor 1 P IHC-P, WB Rb Hu, Mm, Rt, C, D ab23703 Cannabinoid Receptor I P ICC, WB Rb Hu, Rt ab3558 Cannabinoid Receptor I P ICC, IHC-P Rb Hu ab13166 Cannabinoid Receptor I P IF, IHC-Fr, WB Rb Hu, Mm, Rt ab10573 Cannabinoid Receptor II P ICC, IHC-Fr, WB Rb Hu ab3560 Cannabinoid Receptor II P ICC Rb Hu, Rt ab3561

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Dopamine Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

DARPP32 P ELISA, IHC-P, WB Rb Hu, Mm, Rt ab18551 DARPP32 (phospho T34) P ELISA, WB Rb Hu, Rt ab18553 DARPP32 (phospho T75) P ELISA, IHC, WB Rb Hu, Rt ab18552 Dopamine Receptor D1 P ICC, IHC-P Rb Hu ab12969 Neurotransmission Dopamine Receptor D4 P IHC-P Rb Hu ab13318 Neurotensin Receptor 1 P IHC-P Rb Hu ab13201 Neurotensin Receptor 2 P IHC-P Rb Hu ab13068

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Glutamate

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

GRM8 P IHC-P Rb Hu ab13194 Homer (1a+1b+1c) P IHC-Fr, WB Rt Hu, Mm, Rt ab11158 Homer (1b+1c) P ELISA, FACS, ICC, IF, IHC-Fr, IP, RIA, WB Rt Hu, Mm, Rt ab18550 Metabotropic Glutamate Receptor 1 P IHC-P Rb Hu ab13037 Metabotropic Glutamate Receptor 1a P IF, IHC, IHC-Fr, IP, WB Rb F, Mm, Rt ab6439 Metabotropic Glutamate Receptor 1a P ICC, IHC-Fr, WB Rb Hu, Mm, Rt, F ab10302 Metabotropic Glutamate Receptor 2 P WB Rb Rt ab10304 Metabotropic Glutamate Receptor 2 P ICC, IHC-P Rb Hu ab13192 Metabotropic Glutamate Receptor 2 [mG2Na-s] M IF, IHC, WB Mm Rt, Mm ab15672 Metabotropic Glutamate Receptor 2+3 P IHC, IP, WB Rb F, Mm, Rt ab6438 Metabotropic Glutamate Receptor 2+3 P ICC, IHC-Fr, WB Rb Rt ab10307 Metabotropic Glutamate Receptor 3 P WB Rb Rt ab10309 Metabotropic Glutamate Receptor 5 P IHC, IP, WB Rb F, Mm, Rt ab6436 Metabotropic Glutamate Receptor 5+1 P IHC-Fr, IP, WB Rb Mm, Rt ab16214 Metabotropic Glutamate Receptor 6 P WB Rb Rt ab10314 Metabotropic Glutamate Receptor 7 P IHC-P Rb Hu ab13042 mGluR5 P WB Rb Mm, Rt ab10311

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Neuroscience > Neurotransmission > Receptors / Channels > GPCR > More GPCR

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Adenosine A3 Receptor P IHC-P Rb Hu ab13160 APJ Receptor P ICC, IHC-P Rb Hu ab13267 APJ Receptor P IHC-Fr Ch Mm, Rt ab15639 AVPR1B P IHC-P Rb Hu ab13147 AVPR2 P IHC-P Rb Hu ab13149 Calcitonin receptor P ELISA, IHC-P, WB Rb Hu, Mm, Rt ab11042 CCKBR P IHC-P Rb Hu ab13173 CCKBR P WB Rb Hu ab14439 Corticotropin Releasing Factor Receptor 2 P IF, IHC-Fr, IHC-P, WB Rb Hu, Rt ab12964 CysLT1 P IHC-P Rb Hu ab12968 CysLT2 P IHC-P Rb Hu ab13174 EDG5 P WB Rb Hu ab12259 EDG5 P ICC, IHC-PRb Hu ab13127 EDG7 P ICC, IF, WB Rb Hu, Mm, Rt ab23692 EDG7 P IHC-P Rb Hu ab13350 GABA B Receptor 1 P WB Rb Hu ab1135 GABA B Receptor 2 P WB Rb Hu ab1134 GABA B Receptor 2 (phospho S892) P IF, IHC, WB Rb Mm, Rt ab3900 GALR2 P IHC-P Rb Hu ab13179 GALR3 P IHC-P Rb Hu ab13003

Gastric Inhibitory Polypeptide Receptor P IHC-P Rb Hu ab13005 Neurotransmission GHRHR P IHC-P Rb Hu ab13342 GLP1R P IHC-P, WB Rb Hu, Mm, Rt, Ha ab13181 HRH3 P ICC, IHC-PRb Hu ab13014 HRH4 P IHC-P Rb Hu ab13183 LPHN2 P IHC-P Rb Hu ab13019 LPHN3 P IHC-P Rb Hu ab13022 MASS1 P IHC-P Rb Hu ab13150 MC4 Receptor P ICC, IHC-Fr, IHC-P Rb Hu ab13034 Pep. avail. MC4 Receptor P IHC-P, WB Rb Hu, Mm, Rt ab24233 Melatonin Receptor 1A P IHC-P Rb Hu ab13035 Melatonin Receptor 1B P IHC-P Rb Hu ab13357 Melatonin Related Receptor P IHC-P Rb Hu ab13190 Neurokinin A Receptor P ICC, IHC-P Rb Hu ab13397 NMUR1 P IHC-P Rb Hu ab13365 NMUR2 P IHC-P Rb Hu ab13366 NPFF1 Receptor P IHC-P Rb Hu ab13367 NPFF1 Receptor P ELISA, WB Rb Hu ab1401 NPFF2 Receptor P IHC-P Rb Hu ab13198 Pep. avail. NPFF2 Receptor P ELISA, WB Rb Hu ab1402 NPY5R P ICC, IHC-PRb Hu ab13199 OA1 P IHC-P Rb Hu ab13070 OR10R2 P IHC-P Rb Hu ab13072 OR2H3 P IHC-P Rb Hu ab13073 P2Y1 P IHC-P Rb Hu ab13271 P2Y10 P IHC-P Rb Hu ab13272 P2Y11 P IHC-P Rb Hu ab13104 P2Y12 P ICC, IHC-PRb Hu ab13077 Pep. avail. P2Y2 P IHC-Fr Rb Hu, Mm, Rt ab10270 Pep. avail. P2Y8 P IHC-P Rb Hu ab13273 P2Y9 P IHC-P Rb Hu ab13351 Parathyroid Hormone Receptor 2 P IHC-P Rb Hu ab13080 Somatostatin Receptor 1 P IP, WB Rb Hu, Mm, Rt ab479 Somatostatin Receptor 1 P FACS, WB Rb Hu, Rt ab5827 Somatostatin Receptor 2 P IF, WB Rb Hu, Mm, Rt ab9550 Somatostatin Receptor 2 P IF, IHC-P Rb Hu, Rt ab13120 TA4 P IHC-P Rb Hu ab13403

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Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Muscarinic Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Muscarinic Acetylcholine Receptor 1 P WB Rb Mm ab3480 Muscarinic Acetylcholine Receptor 2 P IHC-P Rb Hu ab13043 Muscarinic Acetylcholine Receptor 2 [31-1D1] M IP, WB Mm Hu, Rt, Mm, Pi. Does not react with Ch ab2805 Muscarinic Acetylcholine Receptor M3 P IHC-P Rb Hu ab13063 Muscarinic Acetylcholine Receptor M4 P IHC-P Rb Hu ab13065

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Opioid Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Delta Opioid Receptor P IHC-Fr Rb Mm, Rt, Primate ab10271 Delta Opioid Receptor P ICC Rb Mm, Rt ab10272 Kappa Opioid Receptor P ICC, IHC-Fr Rb Hu ab10283 Kappa Opioid Receptor P WB Rb Mm ab10566 Mu Opioid Receptor P IHC-P Rb Hu ab13074 Mu Opioid Receptor P ICC, IHC-Fr, WB Rb Hu, Mm, Rt, Mk ab10275 Mu Opioid Receptor P IP Rb Mm ab10582 Mu Opioid Receptor splice variant P IHC-Fr Rb Mm, Rt ab10306 Mu Opioid Receptor splice variant P ICC, IHC-Fr Gp Mm, Rt ab10273 Mu Opioid Receptor peptide ab10704

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Serotonin Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/... Neurotransmission

5HT1B Receptor P WB Rb Hu, Mm, Rt ab13896 5-HT1B Receptor P IHC-P Rb Hu ab13154 5HT1D Receptor P WB Rb Hu, Mm ab10132 5HT1D Receptor P WB Rb Hu, Mm, Rt ab13895 5-HT1E Receptor P IHC-P Rb Hu ab13155 5HT1F Receptor P IHC-P Rb Hu ab13156 5HT2B Receptor P IHC-P Rb Hu, Rt ab13292 5HT3 Receptor P IHC Rb Mm, Rt ab16788 5HT3 Receptor P WB Rb Hu, Rt ab13897 5HT4 Receptor P WB Rb Hu, Mm ab10095 5HT6 Receptor P IHC-P Rb Hu ab13293 5HT7 Receptor P IHC-Fr, IHC-P Rb Hu ab13294 5HT7 Receptor P WB Rb Hu, Mm, Rt ab13898 Ketanserin M ELISA Mm Fast trackab1061

5HT2A Receptor antibody (ab16028)

IHC-FrFl - ab16028 staining 5HT2AR on rat brain tissue. Staining seen in coronal section through hypothalamus. Nuclear staining was observed in the Arcuate nucleus. Nuclear-like staining in the nucleus of the solitary tract, but not in the Ventromedial nucleus of the hypothalamus.

Clonality Applications tested Host Species reactivity Datasheet P IHC-FrFl, WB Rb Rt www.abcam.com/ab16028

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Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > AMPA / Kainate

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Glutamate Receptor 1 (AMPA subtype) (phospho S831) P WB Rb Mm, Rt ab3902 Glutamate Receptor 1 (AMPA subtype) (phospho T840) P IP, WB Rb Mm ab12108 Glutamate Receptor 1 (AMPA subtype) (phospho S845) P WB Rb Mm, Rt ab3901 KA1 P WB Rb Hu, Mmab10101

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > GABA Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

GABA A Receptor alpha 1 P IP, WB Rb Rt ab5781 GABA A Receptor alpha 2 P WB Rb Mm, Rt ab8342 GABA A Receptor alpha 4 P IP, WB Rb Mm, Rt ab4120 GABA A Receptor alpha 4 P Dot, WB Rb Rt ab13872 GABA A Receptor alpha 5 P WB Rb Rt, Mm ab3700 GABA A Receptor alpha 5 P WB Rb Hu, Mm ab10098

GABA A Receptor alpha 6 P IP, WB Rb Mm, Rt ab5365 Neurotransmission GABA A Receptor beta 1 P IP, WB Rb Mm, Rt ab16703 GABA A Receptor beta 1 P WB Rb Hu ab23338 GABA A Receptor beta 2 P IP, WB Rb Mm, Rt ab8340 GABA A Receptor beta 3 P IP, WB Rb Mm, Rt ab4046 GABA A Receptor delta P WB Rb Hu, Mm ab10100 Pep. avail. GABA A Receptor gamma 2 P IP, WB Rb Mm, Rt ab4073 GABA A Receptor gamma 3 P WB Rb Hu, Mm, Rt ab13861 Pep. avail. GABA B Receptor 1 P WB Rb Hu ab1135 Pep. avail. GABA B Receptor 2 P WB Rb Hu ab1134 GABA B Receptor 2 (phospho S892) P IF, IHC, WB Rb Mm, Rt ab3900 GABARAP P WB Rb Hu ab1133

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > Glycine Receptor

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Glycine Receptor alpha 1 P IP, WB Rb Hu ab475 Glycine Receptor alpha 1 + alpha 2 P IHC-Fr, WB Rb Rt ab23809 Pep. avail.

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > More Channels

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/... alpha 1 Glycine Receptor P IP, WB Rb ab475

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > nAch Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Nicotinic Acetylcholine Receptor alpha [D6] M FACS, IHC-Fr, IP, WB Mm Hu, Mm ab11149 Nicotinic Acetylcholine Receptor [88B] M IHC-Fr, IP, WB Mm Rt, Mm, Ch, Fi, Am ab2804 Nicotinic Acetylcholine Receptor beta [B3] M FACS, IHC-Fr, IP, WB Mm Hu. Does not react with Mm, Rt or Ch ab11150

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Nicotinic Acetylcholine Receptor gamma [C9] M FACS, IHC-Fr, IP, WB Mm Foetal AChR. Does not react with Adult Hu, Mm, Rt or Ch ab11151 Rapsyn [1234] M ICC, IF, IHC-Fr, WB Mm Mm, Rt, Am, Ch, Fi ab11423 Pep. avail. Syntrophin [1351] M IF, IP, WB Mm Hu, Mm, Rt, Am, Ch, D, Fi ab11425 Syntrophin gamma 2 P ELISA, WB Gt Hu ab15713

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > NMDA Receptors

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

NMDAR1 [R1JHL ] M WB Mm Rt ab1880 NMDAR1 C1 P IHC, IP, WB Rb Rt ab6484 NMDAR1 a/b/c/d P IHC, IP, WB Rb Rt ab6485 NMDAR1 e/f/g/h P IHC, IP, WB Rb Rt ab6486 NMDAR1 N1 P IHC, IP, WB Rb Rt ab6483 NMDAR2A P IHC, IP, WB Rb Mm, Rt, Mk ab14596 NMDAR2B P IP, WBRb Mm ab14400 NMDAR2B (phospho Y1336) P WB Rb Hu, Rt ab14965 NMDAR2B (phospho Y1472) P Dot, WB Rb Hu, Mm, Rt ab16409 NMDAR2C P IHC, IP, WB Rb Hu, Mm, Rt ab110 NMDAR3A + 3B M WB Mm Hu ab2639

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > P2X Receptors

Target Clonality Applications tested Host Species reactivity Datasheet Neurotransmission www.abcam.com/...

P2X1 P ICC, IHC-FrRb Rt ab10248 P2X2 P ICC, IHC-Fr, WB Gp Hu, Rt ab10262 P2X2 P ICC, IHC-Fr, WB Rb Hu, Rt, Mk ab10266 P2X3 P ICC, IHC-Fr, WB Rb Hu, Rt, Mk ab10269

Neuroscience > Neurotransmission > Receptors / Channels > More Ion Channels

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

ARA9 [35-2] M WB Mm Hu, Mm ab468 ASIC beta P IHC-Fr Gp Rt ab10355 DDR1 P ELISA, WB Rb Hu ab5508 DDR2 P ELISA, WBRb Hu ab5520 ENSA P WB Ch Hu, Mm, Rt, B ab14297 GJB1 P IHC-Fr, WB Rb Hu, Rt ab11367 GJB1 [CXN-32] M ELISA, IHC-Fr, WB Mm Hu, Mm, Rt ab11366 SLC31A1 P IHC-P Rb Hu ab13828 TRAR4 P IHC-P Rb Hu ab13139 TRPM7 P WB Gt Hu ab729 TRPM7 P IP Rb Hu. Does not react with Mm or Rt ab16576 Vanilloid Receptor 1 P IF, IHC-Fr, IHC-P Rb Rt ab6166 Vanilloid Receptor 1 P ICC, IHC-Fr Rb Rt ab3486 Vanilloid Receptor 1 P IHC-Fr Gp Rt ab10295 Pep. avail. Vanilloid Receptor 1 P ICC, IHC-Fr, WB Rb Mm, Rt, Mk ab10296 Pep. avail. VRL1 P Dot, IHC-Fr, WB Rb Rt ab6183

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Neuroscience > Neurotransmission > Receptors / Channels > Potassium Channels

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

HCN1 P WB Rb Rt ab19405 KChIP2 P WB Rb Hu, Rt ab3473 KCNQ3 P ICC, IF, IHC-Fr Rb Hu, Mm, Rt ab16228 KCNQ5 P WB Rb Rt ab19319 Pep. avail. Kv beta 2 P ICC, WB Rb Mm ab10665 Kv12 P ICC, WB Rb Mm, Rt ab10667 Kv12 [0T76] M IHC, IP, WB Mm Mm, Rt, Mk ab21091 Kv14 P IF, IP, WB Rb Hu ab16718 Kv22 - Azide free P IHC, WB Rb Rt, X ab21909 Kv42 P IF, IP, WB Rb Rt ab16719

Neuroscience > Neurotransmission > Receptors / Channels > Sodium Channels

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

APXL P Fast track Gt Fast track ab5948 ASIC3 P ICC, IHC-Fr Gp Rt ab10354

Neuroscience > Neurotransmission > Receptors / Channels > Tyrosine Kinase Receptors Neurotransmission Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

EphA1 P ELISA, WBRb Hu ab5385 EphA1 P I-ELISA, WB Gt Hu ab7036 EphA2 P ELISA, WB Rb Ha, Hu, Mm ab5387 EphA2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610 EphA3 P I-ELISA, WB Gt Mm ab7038 EphA3 P ELISA, WB Gt Hu. Does not react with Mm ab10611 EphA3 [III-A4] M FACS, IP Mm Hu ab19439 EphA4 P ELISA, WBRb Hu ab5389 EphA4 P I-ELISA, WB Gt Mm ab7039 EphA4 P I-ELISA, WB Gt Hu ab7041 EphA5 P ELISA, WB Rb Hu, Mm ab5397 EphA5 P ELISA, WB Gt Mm, Rt ab10612 EphA6 P ELISA, WB Gt Mm ab10613 EphA7 P ELISA, WB Rb Hu ab5411 EphA7 P ELISA, WBGt Mm ab10614 EphA8 P ELISA, WBGt Mm ab10615 EphB1 P ELISA, WB Rb Hu, Mm ab5414 EphB1 P I-ELISA, WBGt Rt ab7042 EphB1 P Dot, IP Sh Hu ab10406 EphB2 P ELISA, WB Rb Hu, Mm ab5418 EphB2 P Dot, ELISAGt Mm ab10616 EphB3 P ELISA, WB Gt Mm ab10617 EphB3 P ELISA, WBGt Hu ab7044 EphB4 P Dot, ELISA, Inhib Gt Mm ab10618 EphB6 P Dot, ELISAGt Mm ab10619 Ephrin A2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610 Ephrin A3 P ELISA, WB Gt Hu. Does not react with Mm ab10611 Ephrin A4 P I-ELISA, WB Gt Hu ab7041 Ephrin B3 P ELISA, WB Gt Hu ab7044

Neuroscience > Neurotransmission > Secretory Vesicles > Granins

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Secretogranin II P IHC, IP, WB Rb Hu, Rt ab12241

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Neuroscience > Neurotransmission > Secretory Vesicles > Munc18

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Mint1 P WB Rb Rt ab3448 Munc18 P IP, WB Rb Rt ab3451 MYRIP P Fast track Gt Fast track ab10149 Synaptotagmin P ICC, IP, WB Rb Rt, X ab13260 Synaptotagmin P IF, WB Rb Hu, Rt ab10104 Synaptotagmin P ELISA, WB Ch Hu, B, Mm, Pi, Rt ab8037 Synaptotagmin [ASV30] M ICC, IHC, IP, WB Mm Mm, Rt, X, B, Ch, Rb, Sha ab13259

Neuroscience > Neurotransmission > Secretory Vesicles > Related targets

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Amisyn P Fast track Gt Fast track ab5899 Calpastatin P ELISA, WB Ch Hu ab16423 Calpastatin [2G11D6] M ICC, WB Mm Hu, Rt, C, Pi ab3515 Clathrin light chain [2Q2211] M WB Mm B ab16428 Clathrin light chain [3F133] M IF, IHC-P, IP, WB Mm C ab14408 Clathrin light chain [3F133] - BSA and Azide free M ICC, IF, IHC-P, IP, WB Mm Hu, C ab14409 Pep. avail. Clathrin light chain [CON1] M AP, ICC, IHC, WB Mm Hu, Mm, Rt, C, Sc ab11332 Pep. avail. Clathrin light chain [SPM174], prediluted M IHC-P Mm Hu, C ab17949 Mint1 P WB Rb Rt ab3448 Mint3 P WB Rb Rt, Mmab3450 MYRIP P Fast track Gt Fast track ab10149 SCAMP1 P IP, WB Rb Rt, Pi ab3430 SCAMP2 P WB Rb Hu, Mm, Ha ab3431 SCAMP5 P WB Rb Rt ab3432 Secretogranin II P IHC, IP, WB Rb Hu, Rt ab12241 Neurotransmission SLC6A4 [4A22] M FACS, IHC-P Mm Hu, Rt ab1125 Synaptotagmin P ICC, IP, WB Rb Rt, X ab13260 Synaptotagmin P IF, WB Rb Hu, Rt ab10104 Synaptotagmin P ELISA, WB Ch Hu, B, Mm, Pi, Rt ab8037 Synaptotagmin [ASV30] M ICC, IHC, IP, WB Mm Mm, Rt, X, B, Ch, Rb, Sha ab13259 Synaptotagmin [ASV48] M ICC, IHC, IP, WB Mm Rt, B, Mk, Sha, Mm ab12255

Neuroscience > Neurotransmission > Secretory Vesicles > Rabs

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Rab5 P ICC, IHC, IP, WB Rb Hu, Mm, Rt, B, D, Gp, Ha, Mk ab13253 Rabphilin 3A P WB Rb Rt ab3338 Rabphilin 3A P WB Rb Mm, Rt, B ab12234

Neuroscience > Neurotransmission > Secretory Vesicles > SNAPs & SNAREs

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Amisyn P Fast track Gt Fast track ab5899 SNAP23 P IF, WB Rb Hu, Rt ab4114 SNAP23 P WB Rb Rt, Mm ab3340 SNAP25 P IF, WB Rb Mm, Rt, B ab5666 Pep. avail. SNAP25 [4H251] M WB Mm Hu, Mm, Rt, X, B, Ha, In (tick), Pi ab18002 Pep. avail. SNAP25 [SP12] M ELISA, IHC-P, WB Mm Hu, Ha, Rt, Pi ab11102 Synapsin I (phospho S603) P Dot, WB Rb Rt ab13879 Synapsin I (phospho S9) P WB Rb Hu, Mm, Rt, X , B ab16554

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Synapsin I (phospho S9) P WB Rb Hu ab18006 Synapsin I P AF, IF, IHC-F, IHC-Fr, IP, WB Rb B, Gp, Hu, Mm, Rt ab8 Pep. avail. Synapsin I P IP, WB Rb Hu, Mm, Rt, B ab12239 Pep. avail. Synapsin I P WB Rb Hu, Rt ab18004 Synaptobrevin [4E240] M WB Mm Hu, Mm, Rt, Rb, X, B, Gp, Ha, Mk, Pi, Sh ab18013 Synaptobrevin [4H302] M WB Mm Hu, Mm, Rt, X, B, Ha, Pi ab18015 Synaptobrevin [SP10] M WB Mm Hu, Rt, Ha, Pi ab11109 Synaptobrevin [SP11] M ELISA, IHC-P, WB Mm Hu, Pi. Does not react with Rt ab11104 Synaptophysin P IHC-F, IHC-Fr, IHC-P Rb Hu ab7837 Synaptophysin P IHC-Fr, IP, WB Rb Hu, Mm, Rt ab14692 Synaptophysin [4E206] M IHC, IP, WB Mm Hu, Mm, Rt, B ab18008 Synaptophysin [4H255] M WB Mm Hu, Mm, Rt, B ab18007 Synaptophysin [SP11] M IHC-P, WB Rb Hu ab16659 Synaptophysin [SP15] M ELISA, IHC-P, WB Mm Hu, Rt, Ha, Pi ab11105 Synaptophysin [SVP38] M FACS, ICC, WB Mm Rt ab12353 Synaptophysin [SY38] M IHC-Fr, IHC-P Mm Rt, Mm, B ab8049 Synaptophysin [SYP02] M IHC-Fr, IHC-P Mm Hu ab6245 Synaptophysin, prediluted P IHC-F, IHC-P Rb Hu ab8547 Pep. avail.

Syntaxin 3 P IF, WB Rb Ma ab4113 Neurotransmission Syntaxin P ELISA, IP, WB Ch Hu, B, Mm, Pi, Rt ab8038 Syntaxin P WB Rb Mm, Rt, C ab13262 Syntaxin [1B663] M IP, WB Mm Rt ab18009 Syntaxin [4H256] M WB Mm Hu, Mm, Rt, X, B, F, Ha, Pi ab18010 Syntaxin [SP6] M IHC-Fr, IP, WB Mm Hu, Mm, Rt, X, B, F, Ha, Pi ab12236 Syntaxin [SP8] M ELISA, IHC-Fr, WB Mm Hu, Rt, Pi, Ha ab257 Syntaxin [STX01 (HPC-1)] M IHC-P, WB Mm Hu, Rb, Rt, C. Does not react with Gp ab3265 VAMP8 P IF, IM, IP, WB Rb Hu, Rt, D ab6186

Neuroscience > Neurotransmission > Transporters > Dopamine

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

CSPS P WB Rb Hu, Mm, Rt ab16870 CSPS [3F10] M ELISA, IP Mm Hu, Mm, Rt ab16773 Dopamine Transporter P ELISA, WB Rb Hu, Mm, Rt ab18548 Dopamine Transporter [hDAT-LOOP] M ICC, IHC-Fr, WB Rt Hu, Rt ab5981

Neuroscience > Neurotransmission > Transporters > GABA

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

GABA Transporter 1 / GAT 1 P IF, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch ab426 GABA Transporter 2 P IF, IHC-Fr, WB Rb Hu, Mm, Rt, Ch ab2896 GABA Transporter 3 / GAT 3 P IF, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch ab431

Neuroscience > Neurotransmission > Transporters > Glutamate

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Glutamate Aspartate Transporter P FACS, ICC, IHC, PCC, WB Rb Rt ab416 Glutamate Transporter 1 P FACS, ICC, IF, IHC, PCC, WB Rb Hu, Rt ab417 VGLUT1 P EM, IHC, IP, RIA, WB Rb Hu, Rt ab18106

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

MATH1 P ELISA, IF, IHC-Fr, WB Rb Hu, Mm, Rt ab13483 TBL1 P WB Gt Hu ab2243 Pep. avail.

Neuroscience > Sensory systems > Nociception

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

5 Methoxytryptamine P IHC Rb Rt ab15165 Amitriptyline [1BB831] M ELISA, RIA Mm Fast track ab15176 Amitriptyline [202] - BSA and Azide free M IA Mm Fast track ab20408 ASIC beta P IHC-Fr Gp Rt ab10355 ASIC3 P ICC, IHC-Fr Gp Rt ab10354 Axotrophin P WB Ch Hu, Mm, Rt ab13997 Clonidine P IP Rb Many ab14410 Endorphin alpha neo P IHC-Fr, RIA Rb Rt, Pi ab11140 Neuropeptide FF1 P WB Rb Hu ab3898 NMUR1 P IHC-P Rb Hu ab13365 NMUR2 P IHC-P Rb Hu ab13366 Nociceptin P IHC-Fr Gp Rt, Mk ab10276 Nociceptin P IHC-Fr Rb Rt, Mk ab10277 Nociceptin P ELISA, I-ELISA, IHC-Fr, WB Rb Gp, Rt ab6174 NPFF1 Receptor P IHC-P Rb Hu ab13367 NPFF1 Receptor P ELISA, WB Rb Hu ab1401 NPFF2 P WB Rb Hu ab3899 NPFF2 Receptor P IHC-P Rb Hu ab13198 NPFF2 Receptor P ELISA, WB Rb Hu ab1402

Sensory systems PSD93 P IHC-Fr, WB Rb Rt ab2930 PSD93 P IHC-P, WBGt Mm ab12097 TRPM8 P WB Rb Hu ab3243 Vanilloid Receptor 1 P IHC-Fr Gp Rt ab10295 Vanilloid Receptor 1 P ICC, IHC-Fr, WB Rb Mm, Rt, Mk ab10296 Vanilloid Receptor 1 P IF, IHC-Fr Rb Hu ab3487

SPRR1a antibody (ab18580)

IF - detected in mouse L4 DRG sciatic nerve crush using ab18580 at 1/1000

Clonality Applications tested Host Species reactivity Datasheet P WB, IF Rb Mm. Does not react with Rt www.abcam.com/ab18580

Neuroscience Abwire at www.abcam.com/ neuroscience

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Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

Vanilloid Receptor 1 P IF, IHC-Fr, IHC-P Rb Rt ab6166 VRL1 P Dot, IHC-Fr, WB Rb Rt ab6183

Vanilloid Receptor 1 antibody (ab3486)

ICC - stained cells in dorsal root ganglion cells. ab3486 (1/10,000 for 2h at RT) on adult rat dorsal ganglion cells (cultured for 48hrs and fixed in 4% PFA/15% picric acid).

Clonality Applications tested Host Species reactivity Datasheet P IHC-Fr, ICC Rb Rt www.abcam.com/ab3486

Neuroscience > Sensory systems > Olfaction

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/... Sensory systems

Olfactory Receptor LS189744 P IHC-P Rb Hu ab13071 Olfactory Receptor OR10R2 P IHC-P Rb Hu ab13203 Olfactory Receptor OR2A4 P IHC-P Rb Hu ab13205 Olfactory Receptor OR6N1 P IHC-P Rb Hu ab13204 OR10R2 P IHC-P Rb Hu ab13072 OR2H3 P IHC-P Rb Hu ab13073

Neuroscience > Sensory systems > Touch

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

OA1 P IHC-P Rb Hu ab13070

Neuroscience > Sensory systems > Vision

Target Clonality Applications tested Host Species reactivity Datasheet www.abcam.com/...

APXL P Fast track Gt Fast track ab5948 Arrestin C P WB Ch Hu ab15852 Bestrophin P ICC, IHC, IP, WB Rb Hu, Mm, Rt ab14927 Bestrophin M WB Mm Hu, Pi, Mk ab2182 CHX10 P WB Sh Hu, Mm, Rt, B ab16141 CHX10 P WB Sh Hu, Mm, Rt, B ab16142 Melanopsin P IF, IHC-P, WB Rb Mm, Rt ab19306 Melanopsin P IF, IHC-Fr, IHC-P, WB Rb Rt. Does not cross-react with Mm ab19383 NCKX2 P IHC-Fr, WBRb Rt ab3507 OA1 P IHC-P Rb Hu ab13070 PDE6 alpha P WB Rb C, Sh ab5659 PDE6 alpha P WB Rb Sh ab5660 Peropsin P IHC-P Rb Hu ab13377 RAGE P IHC-Fr, WB Rb Rt, Mm ab3611 RAGE P ELISA, IHC-PGt Hu ab20558 RIT1 P WB Rb Hu ab13322 RIT1 [14G7] M WB Mm Hu, Mm ab16458

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Technical help - Neuroscience protocols and tips CONTENTS Page I. WESTERN BLOTTING (WB) - A BEGINNER’S GUIDE

I. WESTERN BLOTTING (WB) – A BEGINNER’S GUIDE...... 66 Western blotting (or immunoblotting) enables determination of the relative A. Lysate preparation ...... 67 amounts of a specific protein present in different samples by separating B. Preparation of cell lysate from cells in culture ...... 67 proteins on a gel and probing for the specific protein with an antibody C Preparation of cell lysate form tissues ...... 67 targeted to that protein only. D. Determination of the protein concentration levels ...... 67 E. Denaturation and reduction of the protein ...... 68 Here is a beginner’s guide to western blotting and our recommendations F. Electrophoresis ...... 68 when using Abcam antibodies. G. Transfer of proteins onto a membrane ...... 69 H. Blocking the membrane ...... 69 I. Incubation with primary antibody ...... 70 A) Lysate preparation J. Incubation with secondary antibody ...... 70 K. Development methods ...... 70 Lysis buffers L. Determination of the protein size ...... 70 A number of lysis buffers are available and the choice of lysis buffer M. Western blot protocol troubleshooting tips ...... 70 depends on the protein investigated. Where the target protein is tightly N. Stripping ...... 72 bound to cytoskeletal elements, a stronger extraction buffer containing SDS or other detergents is required (see below). II. WESTERN BLOTTING OF PHOSPHO-PROTEINS PROTOCOL . . . 72 Cytoplasmic protein: Use NP-40 or RIPA buffer. III. IMMUNOPRECIPITATION PROTOCOL (IP) ...... 73 Soluble proteins are the easiest proteins to extract from a cell A. Reagents ...... 73 homogenate; thus, this can be done readily even without detergent. B. Immunoprecipitation ...... 73 Membrane-bound protein: Use NP-40 or RIPA buffer. IV. NUCLEAR FRACTIONATION PROTOCOL ...... 73 Transmembrane proteins are contained within membranes, a suitable A. Reagents ...... 73 detergent is essential to extract these proteins. B. Method ...... 74 Nuclear protein: Use RIPA buffer. (see page 67) V. MITOCHONDRIAL PURIFICATION PROTOCOL...... 74 Mitochondrial protein: Use Lysis buffer. (see page 74) VI. SOLUBLE (S-100) MITOCHONDRIAL FRACTIONATION ...... 74 PROTOCOL Whole cell protein extracts: Use NP-40 or RIPA buffer.

VII. DETERMINING IF THE ANTIBODY BINDS ONLY ...... 74 In summary... PHOSPHORYLATED PROTEINS (WB OR IHC) Protein location Buffer recommended VIII. DOT BLOT PROTOCOL ...... 74 Cytoplasmic (soluble) Tris buffer A. Reagents ...... 74 B. Method ...... 74 Cytoplasmic (cytoskeletal bound) Tris-Triton buffer Membrane bound NP-40 or RIPA buffer IX. BLOCKING WITH IMMUNIZING PEPTIDE (BL) PROTOCOL . . . . . 75 Nuclear RIPA buffer X. IMMUNOHISTOCHEMISTRY (IHC-P) – PARAFFIN PROTOCOLS. . . 75 Mitochondrial Use specific protocol A. Antigen retrieval ...... 75 B. ABC immunohistochemical staining – chromogenic methods . . . . 76 Whole Cell NP-40 or RIPA buffer C. ABC immunohistochemical staining – fluorescent methods . . . . . 78 D. IHC troubleshooting tips ...... 78 Cytoskeletal bound proteins Extract Buffer: 10 mM Tris, pH 7.4 XI. IMMUNOHISTOCHEMISTRY (IHC-FR) - FROZEN SECTIONS . . . 79 100 mM NaCl Neuroscience protocols and tips 1 mM EDTA XII. IMMUNOCYTOCHEMISTRY (ICC) PROTOCOL ...... 79 1 mM EGTA 1 mM NaF XIII. DOUBLE IMMUNOFLUORESCENCE (IF) STAINING ...... 80 20 mM Na4P2O7 – SIMULTANEOUS PROTOCOL 2 mM Na3VO4 1% Triton X-100 XIV. DOUBLE IMMUNOFLUORESCENCE (IF) STAINING ...... 80 10% glycerol – SEQUENTIAL PROTOCOL 0.1% SDS 0.5% deoxycholate XV. INDIRECT FLOW CYTOMETRY (FACS) PROTOCOL ...... 81 Soluble protein buffer: XVI. DIRECT FLOW CYTOMETRY (FACS) PROTOCOL ...... 81 20 mM Tris-HCl, pH 7.5 1 mM EGTA (Ca2+ chelator) XVII. DIRECT ELISA PROTOCOL ...... 82 If in doubt, start by using RIPA buffer as it is the “strongest”. XVIII. INDIRECT ELISA PROTOCOL ...... 82

XIX. ELISA USING FLUORESCENT SUBSTRATE ...... 83

XX. CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL . . . . 83

XXI. BUFFERS AND STOCK SOLUTIONS ...... 83

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 66 Neuroscience resources: www.abcam.com/neuroscience RIPA buffer (RadioImmunoPrecipitation Assay) buffer: Final concentrations of inhibitors to use: RIPA buffer contains the ionic detergent sodium deoxycholate as an active Final constituent and is particularly used for nuclear membrane disruption for Protease Stock Inhibitor concentration in nuclear extracts. RIPA buffer gives low background but can denature inhibited (store at -20°C) kinases. It can also disrupt protein-protein interactions (and may therefore lysis buffer be problematic for immunoprecipitations/pull down assays). Trypin, Dilute in water. Aprotinin chynotrypin, 2 µg/ml Do not re-use 50mM Tris HCl pH 8 plasmin when defrosted. 150 mM NaCl 1% NP-40 Dilute in water. 0.5% sodium Deoxycholate Leupeptin Lysosomal 5-10 µg/ml Do not re-use 0.1% SDS when defrosted.

The 10% sodium deoxycholate stock solution (5 g into 50 ml) must be Dilute in water. protected from light. Antipain Trypin, plasmin 2-10 µg/ml Do not re-use The 100 mM EDTA stock solution is made with 1.86 g into 40 ml H2O and when defrosted. then add NaOH to dissolve and adjust pH to 7.4. Finally, adjust the total volume to 50 ml). Dilute in water. Store the buffer at 4°C. Pepstatin A aspartyl 1 µg/ml Do not re-use when defrosted. Nonidet-P40 (NP-40) buffer: Dilute in water. 20 mM Tris HCl pH 8 Serine/Threonine Na-Fluoride 5-10 mM Do not re-use 137 mM NaCl phosphatase 10% glycerol when defrosted. 1% nonidet P-40 Dilute in water. 2 mM EDTA Tyrosine Orthovanadate 1 mM Do not re-use phosphatase Protease inhibitors when defrosted. As soon as lysis occurs, proteolysis, dephosphorylation and denaturation Dilute in ethanol, begin. These events can be slowed down tremendously if samples are PMSF Serine, cysteine 1 mM you can re-use kept on ice or at 4°C at all times and appropriate inhibitors are added the same aliquot. fresh to the lysis buffer.

EDTA chelated calcium and magnesium ions. The buffer (with inhibitors) should be ice-cold prior to Sodium orthovanadate is an inhibitor of all tyrosine protein phosphatases. homogenization.

It must be prepared correctly (see below). Neuroscience protocols and tips Sodium fluoride is an inhibitor of serine/threonine protein phosphatases. Aprotinin is a protease inhibitor. B) Preparation of cell lysate from cells in culture Place the cell culture dish in ice and wash the cells with ice-cold PBS, Leupeptin is a lysosomal protease inhibitor. 7 drain the PBS, then add ice-cold lysis buffer (1ml per 10 cells/100mm Anti-pain inhibits trypsin and plasmin proteases. 2 6 2 dish/150cm flask; 0.5ml per 5x10 cells/60mm dish/75cm flask). Scrape Pepstatin is a reversible inhibitor of aspartic proteases. adherent cells off the dish using a cold plastic cell scraper then gently PMSF is a protease inhibitor. transfer the cell suspension into a pre-cooled microfuge tube. Ready to use cocktails of inhibitors from well known suppliers are often Maintain constant agitation for 30 minutes at 4°C. used but you can make your own cocktail. Centrifuge in a microcentrifuge at 4°C. You may have to vary the Sodium orthovanadate preparation: centrifugation force and time depending on the cell type, a guideline is 20 This needs to be done under the fume hood min at 12000 rpm but this must be determined by the end-user (e.g. leukocytes need a very light centrifugation). • Prepare a 100 mM solution in double distilled water • Set pH to 9.0 with HCl Gently remove the tubes from the centrifuge and place on ice, aspirate the • Boil until colorless supernatant and place in a fresh tube kept on ice, and discard the pellet. • Cool to room temperature • Set pH to 9.0 again • Boil again until colorless C) Preparation of cell lysate from tissues • Repeat this cycle until the solution remains at pH 9.0 after boiling and Dissect the tissue of interest with clean tools, on ice preferably, and as cooling quickly as possible to prevent degradation by proteases. Place the tissue • Bring up to the initial volume with water in round bottom microfuge tubes and immerse in liquid nitrogen to “snap • Store in aliquots at - 20°C freeze”. Store samples at -80°C for later use or keep on ice for immediate homogenization. Note: do not permit great changes in volume during boiling; put a loose lid on the container to protect from evaporation. For a ~5 mg piece of tissue, add ~300 µl lysis buffer rapidly to the tube, Discard if the samples turn yellow. homogenize with an electric homogenizer, rinse the blade twice with another 2x300 µl lysis buffer, then maintain constant agitation for 2 hours Remember to add phosphatase inhibitors to cocktails at 4°C (e.g place on an orbital shaker in the fridge). Volumes of lysis buffer bought when investigating phosphorylation events. must be determined in relation to the amount of tissue present (protein extract should not be too diluted to avoid loss of protein and large volumes of samples to be loaded onto gels, the minimum concentration is 0.1 mg/ml, optimal concentration is 1-5 mg/ml).

Centrifuge for 20 min at 12000 rpm at 4°C in a microcentrifuge. Gently remove the tubes from the centrifuge and place in ice, aspirate the supernatant and place in a fresh tube kept on ice, discard the pellet.

D) Determination of the protein concentration levels Perform a Bradford assay, a Lowry assay or a BCA assay. Bovine serum albumin (BSA) is a frequently used protein standard.

Customer Service Tel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM) 67 Once you have determined the concentration in each sample, you can proteins is used for finger-printing, and when properly constructed can be then freeze them at -20°C or -80°C for later use or proceed to the extremely accurate in resolving all of the proteins present within a cell immunoprecipitation stage (see page 73). (greater than 1,500 KDa).

Here we will be describing techniques for one dimensional electrophoresis. E) Denaturation and reduction of the protein nd Antibodies typically recognize a small portion of the protein of interest We recommend Gel Electrophoresis, A Practical Approach (2 Edition, BD (termed epitope) and this domain may reside within the 3D conformation of Hames and D Rickwood, The Practical Approach Series, IRL press ) as a the protein. To enable access of the antibody to this portion it is necessary reference for basic understanding of 2D electrophoresis protocols. to unfold the protein, i.e. denature it. i) Preparation of PAGE gels To do so, use a loading buffer with the anionic denaturing detergent sodium When separated on a polyacrylamide gel, the procedure is abbreviated as dodecyl sulfate (SDS), and boil the mixture at 95-100°C for 5 minutes. SDS-PAGE (for Sodium Dodecyl Sulfate PolyAcrylamide Gel Electrophoresis). The technique has become a standard means for When SDS is used with proteins, all of the proteins become negatively molecular weight determination. charged by their attachment to the SDS anions. SDS denatures proteins by “wrapping around” the polypeptide backbone - and SDS binds to Polyacrylamide gels are formed from the polymerization of two proteins fairly specifically in a mass ratio of 1.4:1. In so doing, SDS compounds, acrylamide and N,N-methylene- bis-acrylamide (Bis, for confers a negative charge to the polypeptide in proportion to its length - short). Bis is a cross-linking agent for the gels. The polymerization is i.e., the denatured polypeptides become “rods” of negative charge clouds initiated by the addition of ammonium persulfate along with either DMAP with equal charge or charge densities per unit length. It is usually or TEMED. The gels are neutral, hydrophilic, three-dimensional networks necessary to reduce disulphide bridges in proteins before they adopt the of long hydrocarbons crosslinked by methylene groups. random-coil configuration necessary for separation by size by using ß- The separation of molecules within a gel is determined by the relative size mercaptoethanol or dithiothreitol (DTT). In denaturing SDS-PAGE of the pores formed within the gel. The pore size of a gel is determined by separations, therefore, migration is determined not by intrinsic electrical two factors, the total amount of acrylamide present (designated as%T) and charge of the polypeptide, but by molecular weight. the amount of cross-linker (%C). As the total amount of acrylamide SDS grade is of utmost importance: a protein stained background along increases, the pore size decreases. With cross- linking, 5%C gives the individual gel tracts with indistinct or slightly distinct protein bands are smallest pore size. Any increase or decrease in%C increases the pore indicative of old or poor quality SDS. size. Gels are designated as percent solutions and will have two necessary parameters. The total acrylamide is given as a% (w/v) of the Exceptions: acrylamide plus the bis-acrylamide. Thus, a 7.5%T would indicate that there is a total of 7.5 g of acrylamide and bis per 100 ml of gel. A gel 1) Certain antibodies recognize the native form (i.e. non-denatured form) designated as 7.5%T:5%C would have a total of 7.5% (w/v) acrylamide + of the protein. It is imperative in those circumstances to run a Western Blot bis, and the bis would be 5% of the total (with pure acrylamide composing in non-denaturing conditions. the remaining 2.5%). Gels can be purchased ready-made (the Abcam laboratory uses Nu- TM 2) Certain antibodies only recognize protein in its non-reduced form i.e. in Page gels from Invitrogen) or tailor-made in the laboratory (recipes can an oxidized form (particularly on cysteine residues) and the compound be found in laboratory handbooks). Either way, choose carefully the SDS must be removed from the traditional loading buffer and migration percentage of your gel as this will determine the rate of migration and buffer (non reducing conditions). degree of separation between proteins.

In summary… Rule of thumb: The smaller the size of the protein of interest, the higher Protein State WB condition Loading buffer Migration buffer the percentage of mono/bis. The bigger the size of the protein of interest, the lower the percentage of mono/bis. With SDS & Reduced - Reducing & ß-mercaptoethanol With SDS Denatured Denaturing or DTT Protein size (kDa) Gel percentage (%)

4-40 20 Native = Reduced- Native Non- No SDS No SDS Denaturing 12-45 15

Neuroscience protocols and tips 10-70 12.5 With SDS, No Oxidized - ß-mercaptoethanol, With SDS 15-100 10 Denatured Non-Reducing No DTT 25-200 8

Rule of thumb: Use a reducing and denaturing WB condition unless the datasheet specifies otherwise. Acrylamide is a potent cumulative neurotoxin: wear gloves at all times.

To enable visualization of the migration of proteins it is common to include Place gels in the electrophoresis tank as instructed by the manufacturer in the loading buffer a small anionic dye molecule (e.g., bromophenol and bathe in migration buffer. blue). Since the dye is anionic, and small, it will migrate the fastest of any component in the mixture to be separated and provide a migration front to ii) Use of positive controls monitor the separation progress. A positive control lysate is used to demonstrate that the protocol is efficient Glycerol is also added to the loading buffer to increase the viscosity of the and correct and that the antibody recognizes the target protein which may sample to be loaded and hence maintain the sample at the bottom of the not be present in the experimental samples. well, restricting overflow and uneven gel loading. During protein sample treatment the sample should be mixed by vortexing We strongly recommend the use of a positive control before and after the heating step for best resolution. lysate when setting up a new experiment; this will give you immediate confidence in the protocol.

F) Electrophoresis iii) Use of a molecular weight marker Electrophoresis can be one dimensional (i.e., one plane of separation) or A range of molecular weight markers will enable the determination of the two dimensional. One dimensional electrophoresis is used for most routine protein size (see below) and also to monitor the progress of an protein and nucleic acid separations. Two dimensional separation of electrophoretic run. A range of MW markers are commercially available,

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 68 Neuroscience resources: www.abcam.com/neuroscience we would recommend colored markers for convenience (i.e., each MW i) Visualization of all the separated proteins after migration band size will be of a different color and therefore easily detectable). This visualization of protein at this stage is useful to determine if proteins have migrated uniformly and evenly. iv) Loading the samples onto the gel Use special gel loading tips or a micro-syringe to load the complete a) Coomassie stain sample in a narrow well with limited movement at the bottom of the well, Although the gel support provides some friction to molecular motion, as resulting in high quality data. soon as the power is turned off the separated protein bands will begin to Never overfill wells. This could lead to artefacts. diffuse (they are freely soluble in aqueous solution). To prevent diffusion Load 20-40 µg total protein per mini-gel well. of proteins treat the gel with an acetic acid and methanol solution which The gels will be submerged in migration buffer which normally contains causes almost all proteins to precipitate (become insoluble). To visualize SDS, except in native gel electrophoresis. the fixed proteins use Coomassie Brilliant Blue R-250 (“Coomassie stain”) and then wash the stain out of the gel by incubation in a weak solution of Run the gel for the recommended time as instructed by the manufacturer, acetic acid and methanol. The stain will not bind to the acrylamide, and this can vary from machine to machine (1 hour to overnight). will wash out (leaving a clear gel). However, it remains strongly bound to When the dye molecule (the “migration front”) reaches the bottom of the the proteins in the gel, and these take on a deep blue color. gel, the power is turned off and the experiment halted. Please note: this is not reversible so if you choose to transfer the proteins v) Loading control troubleshooting tips opt for a copper stain method to check migration is adequate. Loading controls are required to check that the lanes in your gel have been evenly loaded with sample, especially when a b) Copper Stain comparison must be made between the expression levels of Soak freshly-electrophoresed gels in 0.3 M CuCl2 for up to 15 min, a protein in different samples. washing the gels briefly in de-ionized water, and view them against a dark- They are also useful to check for even transfer from the gel to the field background. Proteins come up as clear zones in a translucent blue membrane across the whole gel. Where even loading or transfer have not background. occurred, the loading control bands can be used to quantify the protein amounts in each lane. For publication-quality work, use of a loading Gels may be destained completely by repeated washing in 0.1- 0.25 M control is absolutely essential. Tris/0.25 M EDTA pH 8.0, and then electroblotted, or eluted from the gel for other purposes. Sample type Loading Molecular Caution control weight (kD) Visualization of protein at this stage is useful to determine if proteins have migrated uniformly and evenly. Whole cell beta actin 43 Not suitable for skeletal /cytoplasmic muscle samples. Changes in ii) Visualization after transfer onto membrane: Ponceau Red cell-growth conditions and Wash the membrane in TBST. Dilute the stock Ponceau Red 1:10. The interactions with extracellular stock is made of 2% Ponceau S in 30% trichloroacetic acid and 30% matrix components may alter sulfosalicylic acid. Incubate on an agitator for 5 min. Wash extensively in Neuroscience protocols and tips actin protein synthesis (Farmer water until the water is clear and the protein bands are well-defined. et al., 1983). The membrane may be destained completely by repeated washing in GAPDH 30-40 Some physiological factors, TBST or water. When using a PVDF membrane, re-humidify the such as hypoxia and diabetes, membrane with methanol then wash again in incubation buffer (TBST). increase GAPDH expression in certain cell types. TBST (also called TBS) is a Tris Buffered Saline Tween20 buffer. Some tubulin 55 Tubulin expression may vary laboratories use PBST but we recommend a tris based buffer. according to resistance to antimicrobial and antimitotic TBS 10x (concentrated TBS) drugs (Sangrajrang S. et al, 24.23 g Trizma HCl 1998, Prasad V et al, 2000) 80.06 g NaCl Mix in 800 ml ultra pure water. Mitochondrial VDAC1 / 31 pH to 7.6 with pure HCl. Porin Top up to 1 L. COXIV 16 Many proteins run at the same 16 kD size as COXIV. TBST Nuclear Lamin B1 66 Not suitable for samples where For 1 L: 100 ml of TBS 10x + 900 ml ultra pure water + 1ml Tween20 nuclear envelope is removed. Tween20 is very viscous and will stick to the tip of your measuring pipettes Tata binding 38 Not suitable for samples where so make sure you add the right amount of the detergent to the tris buffer. protein TBP DNA is removed. Keep TBST at 4°C to prevent contamination and do not keep it for more than 1 week.

G) Transfer of proteins onto a membrane (WB) Do not add azide to TBST as it will quench the enzyme horse radish Transfer can be done in wet or semi-dry conditions. Wet conditions are peroxidase (HRP) conjugated to the secondary antibody and prevent recommended; especially for high MW proteins (if a semi-dry transfer development of the signal. system is used for large proteins add SDS to help transfer). Use the power conditions recommended by the manufacturer. H) Blocking the membrane Blocking the membrane prevents non-specific background binding of the Two types of membranes are available: nitrocellulose and PVDF. The primary and/or secondary antibodies to the membrane (which has a high choice is personal and both work very well. capacity at binding proteins and therefore antibodies). Two blocking solutions are traditionally used: non-fat milk or BSA. PVDF membranes require careful pre-treatment: cut the membrane to the appropriate size then soak it in methanol for 1-2 min. Incubate in ice cold Milk is cheaper but is not recommended for studies of phosphoproteins transfer buffer for 5 min. (milk contains casein which is a phosphoprotein; this is why it causes high background because the phospho-specific antibody detects the casein The gel needs to equilibrate for 3-5 min in ice cold transfer buffer. Failure present in the milk). to do so will cause shrinking while transferring and an unsightly pattern of transfer.

Customer Service Tel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM) 69 To prepare this solution weigh 5 g per 100 ml of Tris Buffer Saline membrane, transforming the signal into a digital image for rapid analysis Tween20 (TBST, also called TBS) buffer. Mix well and filter. Failure to filter with software provided with the detection machine. can lead to “spotting” where tiny dark grains will contaminate the blot A range of machines are now commercially available. during development. At the front of the next generation are systems which do not use HRP- Incubate for 1 hr at 4°C under agitation. Rinse for 5 sec in TBST after the conjugated antibodies (i.e chemiluminescence): for example, STORM incubation. Analysers detect fluorescence from fluorochrome-conjugated secondary antibodies, the Odyssey Infrared Imaging System detects infrared I) Incubation with primary antibody fluorescence. Incubation Buffer: Dilute the antibody in TBST at the suggested dilution, if the datasheet does not have a recommended dilution try a range of L) Determination of the protein size dilutions (1:100-1:3000) and optimize the dilution according to the results. With SDS treatment, the proteins will migrate as a function of their Too much antibody will result in non-specific bands. molecular mass. It is traditional in certain laboratories to incubate the antibody in blocking A linear relationship therefore exists between the logarithm of the buffer, while other laboratories incubate the antibody in TBST without a molecular weight of an SDS-denatured polypeptide, or native nucleic acid, blocking agent. The results are variable from antibody to antibody and you and its Rf. The Rf is calculated as the ratio of the distance migrated by the may find it makes a difference to either use no blocking agent in the molecule to that migrated by a marker dye-front. A simple way of antibody buffer or the same agent as the blocking buffer. determining relative molecular weight by electrophoresis (Mr or MW) is to plot a standard curve of distance migrated vs. log 10MW for known Incubation Time: The time can vary between a few hours and overnight samples (molecular weight markers), and read off the logMr of the sample (rarely more than 18 hours), and is dependent on the binding affinity of the after measuring the distance migrated on the same gel. antibody for the protein and the abundance of protein. We recommend a more dilute antibody for a prolonged incubation to ensure specific binding. M) Western blot protocol troubleshooting tips

Incubation Temperature: preferably cold. If incubating in blocking buffer, No signal it is imperative to incubate at 4°C or contamination will incur and thus destruction of the protein overnight (especially phospho groups). The primary antibody and the secondary antibody are not compatible. Use secondary antibody that was raised against the species in which the Agitation of the antibody is recommended to enable adequate primary was raised (e.g primary is raised in rabbit, use anti-rabbit homogenous covering of the membrane and prevent uneven binding. secondary).

J) Incubation with secondary antibody Not enough primary or secondary antibody is bound to the protein of Incubation Buffer: Dilute the antibody in TBST at the suggested dilution, interest. if the datasheet does not have a recommended dilution try a range of dilutions (1:1000- 1:20,000) and optimize the dilution according to the Use more concentrated antibody. Incubate longer (e.g. overnight) at 4ºC. results. Too much antibody will result in non-specific bands. As with the primary antibody you may choose to incubate the secondary Cross-reaction between blocking agent and primary or secondary antibody in blocking buffer or not. antibody.

Incubation Time: 1-2 hours. Use a mild detergent such as Tween20 or switch blocking reagent (i.e. commonly used blocking reagents are milk, BSA, serum or gelatin). Incubation Temperature: room temperature. What secondary antibody to choose? We recommend HRP-conjugated The primary antibody does not recognize the protein in the species secondary antibodies. being tested. ALP-conjugated secondary antibodies (alkaline phosphatase) are not recommended as they are not sensitive enough. Check the datasheet or perform a ClustalW alignment to ensure your antibody should react with the target protein; Run the recommended Agitation of the antibody is recommended to enable adequate positive control. homogenous covering of the membrane and prevent uneven binding. Insufficient antigen. K) Development Methods Detection kits Load at least 20-30 ug protein per lane; Use protease inhibitors; Run the Neuroscience protocols and tips For HRP-conjugated antibodies: ECL and ECL+ (home made or recommended positive control. commercially available) are the traditional kits used and we recommend ECL+ or more sensitive new generation kits (especially for the new The protein of interest is not abundantly present in the tissue. generation detection machines such as Genegnome, etc, in those cases use the detection kit recommended by the manufacturer of the machine). Use an enrichment step to maximize the signal (e.g. prepare nuclear We do not recommend ECL or BCIP/NBT detection kits as they are not lysates for a nuclear protein, etc.). very sensitive. Poor transfer of protein to membrane. X-ray films: Manual film development is traditionally used and enables the scientist to Check the transfer with a reversible stain such as Ponceau S; check that control the incubation time of the x-ray film in the developing agent and the transfer was not performed the wrong way; if using PVDF membrane fixation agent. make sure you pre-soak the membrane in MeOH then in transfer buffer.

Automated x-ray film developers are also widely used and easy to use. Excessive washing of the membrane.

Remember that an over-exposed film is not suitable for Do not over wash the membrane. analysis as determination of the relative amount of protein is not possible (overexposed films show totally Too much blocking does not allow you to visualize your protein of black bands with no contrast, numerous non-specific interest. bands, etc…). Instead of using 5% milk in the antibody buffers try removing the milk or Digital images: using 0.5%; Switch blocking reagents or block for less time. The new generation of film developers are light tight units with a camera detecting the minute about of chemiluminescence emanating from the

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 70 Neuroscience resources: www.abcam.com/neuroscience

No signal Multiple bands Over-use of the primary antibody. Unreported novel proteins or different splice variants that share similar epitopes and could possibly be from the same protein family Use fresh antibody as the effective concentration is lowered upon each re-use. are being detected.

Secondary antibody inhibited by sodium azide. Check the literature for other reports and also perform a BLAST search; Use the cell line or tissue reported on the datasheet. Do not use sodium azide together with HRP-conjugated antibodies. Primary antibody concentration is too high - at high concentration Detection kit is old and substrate is inactive. multiple bands are often seen.

Use fresh substrate. Try decreasing the antibody concentration and/or the incubation period.

Secondary antibody concentration is too high - at high concentration High background secondaries will bind non-specifically. Blocking of non-specific binding might be absent or insufficient. Try decreasing the concentration. Run a secondary antibody control Increase the blocking incubation period and consider changing blocking (without the primary). agent. Abcam recommends 5% non-fat dry milk, 3% BSA, or normal serum for 30 min. These can be included in the antibody buffers as well. The antibody has not been purified.

The primary antibody concentration may be too high. Try to use affinity purified antibody. This will often remove non-specific bands.

Titrate the antibody to the optimal concentration, incubate for longer but in The bands may be non-specific. more dilute antibody (a slow but targeted binding is best). Where possible use blocking peptides to differentiate between specific and Incubation temperature may be too high. non-specific bands. Only specific bands should be blocked (and thus disappear). Incubate blot at 4°C. The protein target may form multimers. The secondary antibody may be binding non-specifically or reacting with the blocking reagent. Try boiling in SDS-Page for 10 minutes rather than 5 minutes to disrupt multimers. Run a secondary control without primary antibody.

Cross-reaction between blocking agent and primary or secondary. Uneven white “spots”on the blot Neuroscience protocols and tips Air bubbles were trapped against the membrane during transfer or Add a mild detergent such as Tween20 to the incubation and washing buffer. the antibody is not evenly spread on the membrane.

(phospho-specific protein) Milk contains casein which is a Make sure you remove bubbles when preparing the gel for transfer. phosphoprotein; this is why it causes high background because the Incubate antibodies under agitation. phospho-specific antibody detects the casein present in the milk.

Use BSA as a blocking reagent instead of milk. Black dots on the blot

Washing of unbound antibodies may be insufficient. The antibodies are binding to the blocking agent. Filter the blocking agent. Increase the number of washes.

Your choice of membrane may give high background. White bands on a black blot (negative of expected blot) Nitrocellulose membrane is considered to give less background than PVDF. Too much primary and/or too much secondary antibody. Dilute the antibodies more. The membrane has dried out.

Care should be taken to prevent the membrane from drying out during incubation. MW marker lane is black The antibody is reacting with the MW marker. Multiple bands Add a blank lane between the MW marker and the first sample lane. Cell lines that have been frequently passaged gradually accumulate differences in their protein expression profiles.

Go back to the original non-passaged cell line and run the current and The band of interest is very low/high on the blot original cell line samples in parallel. Separation is not efficient.

The protein sample has multiple modified forms in vivo such as Change the gel percentage: a higher percentage for small protein, lower acetylation, methylation, myristylation, phosphorylation, percentage for large proteins. glycosylation etc.

Examine the literature and use an agent to dephosphorylate, de- Smile effect of the bands glycosylate, etc. the protein to bring it to the correct size. 1. Migration was too fast. 2. Migration was too hot (changing the pH and altering the migration). The target in your protein sample has been digested (more likely if the bands are of lower molecular weight). Slow down the migration or run the gel in the cold room or on ice.

Make sure that you incorporate sufficient protease inhibitors in your sample buffer.

Customer Service Tel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM) 71 Uneven band size in lanes probed for the same protein II. WESTERN BLOTTING OF PHOSPHO-PROTEINS Gel has set too quickly while casting and the acrylamide percentage PROTOCOL is not even along the lanes. Homogenize the cells or tissue of interest in lysis buffer made fresh and Review the recipe of the gel and the addition of TEMED to the gels, add a containing a cocktail of protease inhibitors (and phosphatase inhibitors little 0.1% SDS in water to the top of the migrating gel while it sets to stop when dealing with phosphorylated proteins). it from drying. As soon as lysis occurs, proteolysis, dephosphorylation and denaturation begin. These events can be slowed down tremendously if samples are Uneven staining of the gel kept on ice or at 4°C at all times and appropriate inhibitors are added 1. Contamination from bacteria fresh to the lysis buffer. 2. Not enough antibody Use a RIPA or NP40 buffer supplemented with fresh protease and 1. Keep antibodies at 4˚C and use fresh buffers covers the gel. phosphatase inhibitors: 2. Make sure the membrane is covered with the antibody/ incubate under agitation. Ready to use cocktails of inhibitors from well known suppliers are often used but you can make your own cocktail. N) Stripping Stripping is the term used to describe the removal of primary and Remember to add phosphatase inhibitors to cocktails secondary antibodies from the membrane. This step may remove some bought when investigating phosphorylation events. protein from the membrane or damage fragile protein and a PVDF membrane is highly recommended. Final concentrations of inhibitors: see table on page 67

Stripping is used when more than one protein is investigated on the same blot, 1. To a sample of protein solution containing 1-100 ng of the target protein or the same protein with different antibodies (for example a phospho-specific (500 ug lysate), add an equal volume of 2x SDS-PAGE sample buffer. antibody is probed, then the relative total amount of protein is calculated). For reduced samples, the sample buffer should be supplemented with DTT or ß-mercaptoethanol. For non-reduced samples, the DTT or ß- Three types of stripping are found. As a rule of thumb, try the gentler one mercaptoethanol is not added. first then proceed to harsher ones if you still get signal. Most antibodies require medium or harsh stripping to remove all signal. 2. Denature the proteins by heating the sample to 95˚C, or boiling, for 5 min.

Gentle stripping: 3. Load the sample onto an SDS-polyacrylamide gel and run the gel under 10 min PBS standard conditions. 10 min PBS 20 min TBST 4. Transfer the proteins to a PVDF membrane using semi-dry or wet 20 min TBST transfer methods. Please note: for PVDF it is essential to pre-wet the membrane in methanol prior to transfer. Ready for blocking stage. 5. If required, the efficiency of transfer can be determined by staining the Medium stripping: membrane briefly (10 s) in Ponceau stain. The stain can be removed by Make fresh stripping buffer: washing in PBST or TBST. We would recommend not washing blots in 15 g glycine distilled water as this can strip off proteins in some circumstances. 1 g SDS 10 ml Tween20 6. Block the membrane with 5% w/v BSA in TBST. Incubate for 1 h at Set the pH to 2.2 4°C with agitation.

make up to 1 L with ultrapure water 7. Dilute the primary antibody in TBST to the recommended dilution. We recommend incubating in a sealed bag, hybridization tube or 50 ml Falcon Membrane incubation: tubes (~2.5 ml primary antibody/blot). Incubate overnight at 4˚C with 5-10 min stripping buffer agitation. 5-10 min stripping buffer 10 min PBS 8. Rinse the blot in TBST three to four times for 5 min each at room Neuroscience protocols and tips 10 min PBS temperature. 5 min TBST 1 min methanol rehydration (PVDF membrane only) 9. Dilute the horseradish peroxidase (HRP) labeled secondary antibody at 5 min TBST the recommended dilution (1/5000 is usually a good working dilution although this needs to be optimized for the particular application) in TBST. Ready for blocking stage. 10. Rinse the blot in TBST three to four times for 5 min each at room Harsh stripping: to be done under the fumehood temperature. For 100 ml: 20 ml SDS 10% 11. Perform ECL Plus detection. 12.5 ml Tris HCl pH 6.8 0.5M 67.5 ml ultra pure water Top tips for WB of phosphorylated proteins:

Add 0.8 ml ß-mercaptoethanol under the fumehood. 1. Keep the proteins in their phosphorylated state! Add adequate phosphatase inhibitors and keep samples on ice at all times. Make stripping buffer: 2. Block the membrane in 5% w/v BSA (fractionV) NOT MILK (milk Warm up the buffer at 50°C. contains casein which is a phosphoprotein; This is why it causes Add the buffer to a small plastic box which has a tight lid. high background because the phospho-specific antibody detects the Add the membrane. Incubate at 50°C for up to 45 min. casein present in the milk). Dispose of the solution as required for ß-mercaptoethanol based buffers. 3. Remember the phosphorylation may need to be induced. Low Rinse the membrane under a running water tap for 1-2 hours. signal or no signal may mean that the induction is not sufficient. Run Traces of ß-mercaptoethanol will damage the antibodies. Wash extensively. the recommended positive control with your samples. 1 min methanol rehydration (PVDF membrane only) 5 min TBST

Ready for blocking stage.

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Immunoprecipitation is a method that enables the purification of a protein. Species Immunoglobulin Isotype Protein A Protein G An antibody for the protein of interest is incubated with a cell extract so Human IgG1   that the antibody will bind the protein in solution. The antibody/antigen complex will then itself be pulled out of the sample using protein A/G- IgG2   coupled agarose beads. This physically isolates the protein of interest from the rest of the sample. The sample can then be separated by SDS-PAGE IgG3 -  for WB analysis. IgG4   A) Reagents Lysis buffers used for IP: RIPA buffer, NP40 buffer, i.e buffers with some IgM Use anti Human IgM detergent and with protease inhibitors (and phosphatase inhibitors if IgE  looking at phospho-proteins). - IgA  sterile PBS pH 7.4 - Mouse IgG1   sterile PBS-BSA 1% (filtered) IgG2a   TBST buffer IgG2b   loading/sample buffer used for WB IgG3  

B) Immunoprecipitation: IgM Use anti Mouse IgM 1. On ice, in a tube add 10-500 µg cell lysate plus 0.5-5 µg of antibody. These amounts will be chosen depending on the abundance of the protein Rat IgG1 -  and the affinity of the antibody for the protein, typically in a pilot experiment where a fixed amount of protein is precipitated by increasing IgG2a -  amounts of antibody. IgG2b -  2. Incubate the sample with the antibody between 1 hour to overnight (depending again on the amount of protein and affinity properties of the IgG2c   antibody), at 4°C, preferably under agitation. Chicken All isotypes - 

3. Meanwhile prepare the sepharose beads. If using a monoclonal Neuroscience protocols and tips Cow All isotypes   antibody choose protein G-coupled sepharose beads, if using a polyclonal antibody choose protein A-coupled sepharose beads. If the beads come Goat All isotypes  as a powder incubate 100 mg of beads in 1ml PBS 0.1M, wash for one - hour so they swell up, then centrifuge, remove the supernatant and Guinea Pig All isotypes   discard, add 1ml PBS-BSA 1% w/v, mix for one hour and rinse in PBS twice. Remove the supernatant and add 400 µl of buffer made with Hamster All isotypes   protease inhibitors (can be the same as the lysis buffer). The slurry is now ready for use. It can be stored at 4°C for a few days; for longer periods Horse All isotypes   keep the beads in PBS with 0.02% azide (rinse extensively the beads on the day of use and make up in lysis buffer). You can also buy pre-swollen Pig All isotypes   beads as slurry ready for use. Rabbit All isotypes  

IgM antibody: Do not use protein-A or protein-G Sheep All isotypes  conjugated beads, use Goat anti Mouse IgM (or - polyvalent Ig, or anti-heavy chain) beads.

4. Mix the slurry well and add 70-100 ul of the beads to each sample. Always IV. NUCLEAR FRACTIONATION PROTOCOL keep samples on ice. Beads will tend to stick to the sides of the tip so try to minimize the movement in the pipette and use a tip cut 5 mm from the top. A) Reagents 2 5. Incubate the lysate-beads mixture at 4°C under rotary agitation for 4 Buffer A – 10 mM HEPES, 1.5 mM MgCl , 10 mM KCl, 0.5 mM DTT, hours (the optimal incubation time can be determined in a preliminary 0.05% NP40 (or 0.05% Igepal or Tergitol) pH 7.9 experiment). To prepare 250 ml stock of buffer A – HEPES: 1M = 238.3 g/L, therefore 10 mM = 0.59 g/250ml 6. When the incubation time is over, centrifuge the tubes, remove the MgCl2: 1M = 203.3 g/L, therefore 1.5 mM = 0.076 g/250ml supernatant and wash the beads in lysis buffer three times (each time centrifuging at 4°C and removing the supernatant). KCl: 1M = 74.5 g/L, therefore 10 mM = 0.187 g/250ml DTT: 1M = 154.2 g/L, therefore 0.5 mM = 0.019 g/250ml 7. Finally, remove the last supernatant and add 25-50 µl of 2x loading NP40 = 0.05% buffer. Boil at 95-100°C for 5 minutes to denature the protein and separate it from the protein-A/G beads, then centrifuge and keep the supernatant Buffer B – 5 mM HEPES, 1.5 mM MgCl2, 0.2 mM EDTA, 0.5 mM DTT, where the protein is now. You can then freeze the samples or run them on 26% glycerol (v/v), pH 7.9 a SDS-PAGE. See WB protocol for further details (page 66). To prepare 250 ml stock of buffer B – HEPES: 1M = 238.3 g/L, therefore 5mM = 0.295 g/250ml MgCl2: 1M = 203.3 g/L, therefore 1.5mM = 0.076 g/250ml EDTA: 1M = 372.2 g/L, therefore 0.2mM = 0.0186 g/250ml DTT: 1M = 154.2 g/L, therefore 0.5mM = 0.019 g/250ml 26% Glycerol (v/v) = 65ml

4.6 M NaCl – 87.66 g/326ml

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B) Method 3. Spin the final mitochondrial lysate at 100,000 g in an ultracentrifuge 1. Prepare 1 ml of buffer A with added cocktail of usual inhibitors from using a TY65 Beckman rotor at 4˚C, for 60 minutes. frozen stock and store on ice. 4. Carefully collect the clear supernatant, avoiding the fluffy layer over the 2. Add 500 µl of buffer a per large petri dish on ice and scrape thoroughly, pellet, to yield the final S-100 fraction. leave on ice for 10 min. 5. Freeze in aliquots, in liquid nitrogen, and store at -80˚C. 3. Centrifuge at 4˚C at 3000 rpm for 10 min. This is a slightly modified protocol taken from Methods in Enzymology, Vol 4. Remove supernatant and keep it (this will contain everything except 264, by Vicente Micol, Patricio Fernandez-Silva, and Giuseppe Attardi. large plasma membrane pieces, DNA, nucleoli), extract out 10 µl for Bradford assay. VII. DETERMINING IF THE ANTIBODY BINDS ONLY 5. On ice resuspend pellet in 374 µl of buffer B and add 26 µl of 4.6 M PHOSPHORYLATED PROTEINS (WB OR IHC) PROTOCOL NaCl to give 300mM NaCl (high salt helps lyse membranes and forces DNA into solution). 1. Tissue sections or PVDF membranes (after protein transfer and rehydratation of the membrane in 100% methanol followed by water) 6. Homogenize with 20 full strokes in Dounce or glass homogenizer on ice. should be incubated overnight at 37°C in 1% calf intestinal alkaline phosphatase (CIP, New England Biolabs, Beverly, MA, USA) diluted in NE 7. Leave on ice for 30 min. buffer 1 (in mM: NaCl, 10; Tris, 5, pH 7.9; MgCl2, 1; dithiothreitol, 0.1).

8. Centrifuge at 24,000 g for 20 min at 4˚C. 2. Sister sections or membranes should be incubated in the NE buffer only. Rinse three times in PBST (IHC) or TBST (WB) the sections or membranes, 9. Aliquot supernatant, remove 10 µl for Bradford assay and store at -70˚C. respectively.

3. In PBST (IHC) or TBST (WB), membranes in 5% milk. V. MITOCHONDRIAL PURIFICATION PROTOCOL 4. Incubate sections and membranes overnight in primary antibody. This protocol is for use with cells in suspension. 5. Secondary antibodies concentrations and revelation protocol should be 1. Collect cells by centrifugation at approximately 370 g for 10 minutes. performed as usual. Decant supernatant and re-suspend cells in 10 packed cell volumes of NKM buffer (1 mM TrisHCl, pH 7.4, 0.13 M NaCl, 5 mM KCl, 7.5 mM MgCl2). Ref: European Journal of Neuroscience, (2005) 21 (7) pp. 1785–97, Sophie Pezet, Achillefs Spyropoulos, Robert J. Williams and Stephen B. McMahon. 2. Pellet cells and decant supernatant, repeat this washing step 2 times. Resuspend cells in 6 packed cell volumes of homogenization buffer (10 mM Tris-HCl, pH 6.7, 10 mM KCl, 0.15 mM MgCl2, 1 mM PMSF, and 1 mM DTT, always add PMSF and DTT immediately before use). VIII. DOT BLOT PROTOCOL

3. Transfer cells to a glass homogenizer and incubate for 10 minutes on A technique for detecting, analyzing, and identifying proteins, similar to the ice. Using a tight pestle, homogenize the cells. Check under the western blot technique but differing in that protein samples are not microscope for cell breakage, the optimum is around 60%. This may separated electrophoretically but are spotted through circular templates require 30 strokes or so of the pestle. directly onto the membrane or paper substrate.

4. Pour homogenate into a conical centrifuge tube containing 1 packed cell Concentration of proteins in crude preparations (such as culture volume of 2 M sucrose solution and mixed gently. Pellet unbroken cells, supernatant) can be estimated semi-quantitatively by using “Dot Blot” nuclei, and large debris at 1200 g for 5 minutes and transfer the method if you have both purified protein and specific antibody against it. supernatant to another tube. This treatment is repeated twice, transferring the supernatant to a new tube each time, discarding the pellet. A) Reagents TBS: 20 mM Tris-HCl, 150 mM NaCl, pH 7.5 5. Pellet the mitochondria by centrifuging at 7000 g for 10 minutes. TBS-T: 0.05% Tween20 in TBS

Neuroscience protocols and tips Resuspend the mitochondrial pellet in 3 packed cell volumes of BSA/TBS-T: 0.1% BSA in TBS-T mitochondrial suspension buffer (10 mM TrisHCl. pH 6.7, 0.15 mM MgCl2, Nitrocellulose membrane (BIO-RAD, Trans-Blot, etc.) 0.25 mM sucrose, 1 mM PMSF, 1 mM DTT). Spin at 9500 g for 5 minutes to re-pellet the mitochondria. B) Procedure 1. Have nitrocellulose membrane ready, draw grid by pencil to indicate the 6. At this point, you can add 1X protein gel loading buffer and run on a gel region you are going to blot (see below). if a whole mitochondrial protein extract is needed, further purify the mitochondria on a sucrose gradient if you really need very pure 2. Using narrow-mouth pipet tip, spot 2 µl of samples onto the mitochondria, or purify a soluble (S-100) fraction of the mitochondria. This nitrocellulose membrane at the center of the grid. Minimize the area that last protocol is given below. the solution penetrates (usually 3-4 mm diam.) by applying it slowly.

3. Let the membrane dry. VI. SOLUBLE (S-100) MITOCHONDRIAL FRACTIONATION PROTOCOL 4. Block non-specific sites by soaking in 5% BSA in TBS-T (0.5-1 hr, RT). Use 10cm Petri Dish for reaction chamber. 1. Resuspend mitochondria in one-third the packed cell volume 5. Incubate with primary antibody (0.1-10 µg/ml for purified antibody, mitochondrial lysis buffer (25 mM HEPES-KOH pH 7.6, 5 mM MgCl2, 0.5 1:1000 to 1:100000 dilution for antisera, 1:100 to 1:10000 for hybridoma mM EDTA, 10% glycerol, 1 mM DTT, 1 mM PMSF). Put the suspension supernatant) dissolved in BSA/TBS-T for 30 min at RT. into a glass homogenizer and homogenize with 10 strokes using a tight pestle. Add Tween20 and KCl to final concentrations of 0.5% and 0.5 M, 6. Wash three times with TBS-T (3 x 5 min). respectively.

2.Incubate the mixture on ice 5 minutes. The homogenization is repeated for a total of 10 times.

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7. Incubate with secondary antibody conjugated with HRP (for optimum i) Enzymatic Antigen Retrieval dilution, follow the manufacturer’s recommendation) for 30 min at RT. Tissue sections are best mounted on APES (amino-propyl-tri-ethoxy- silane) coated slides. Slides should be placed in a rack for this procedure. 8. Wash three times with TBS-T (15 min x 1, 5 min x 2), then once with TBS (5 min). Materials • Water baths 9. Incubate with ECL reagent for 1 min, cover with Saran-wrap (remove • 2 troughs excessive solution from the surface), and expose X-ray film in the dark • pH meter room. Try several different lengths of exposure. • Magnetic stirrer • Thermometer 10. Compare the signal from your unknown sample to that of standard and estimate the concentration. Reagents • Alpha-Chymotrypsin (type II from Bovine pancreas) 0.1 g • Calcium Chloride 0.1 g IX. BLOCKING WITH IMMUNIZING PEPTIDE (BL) • Ultra-pure water 100.0 ml • 0.5% sodium hydroxide solution PROTOCOL • 0.5% hydrochloric acid solution • Xylene It is not uncommon to see more than one band in Western blotting when • Industrial methylated spirits (IMS) or methanol probing with a given antibody or to see more diffuse staining in immunolocalization studies. The question arises which band or staining is Method specific. 1. Set water bath to 37°C. Add the required amount of ultra pure water into each trough then place the troughs into the water bath (See note i). Allow The antibody specificity is generally studied by competing with excess the ultra pure water to warm to 37°C. antigen (peptide or protein) or immuno-neutralization with the antigens. 2. De-wax and re-hydrate paraffin sections by placing them in 3 changes In principle, a small volume of antibody (e.g. 1-5 µl) is first reacted with of xylene for 3 minutes each, followed by 3 changes of IMS or methanol excess peptide (5-50 fold over the antibody; e.g. 1 µg antibody reacted for 3 minutes each, followed by cold running tap water for 3 minutes. At no with 5-50 µg peptide; exact amounts determined by titration) to neutralize time from this point onwards should the slides be allowed to dry out! Place it. The neutralized antibody can no longer bind to another antigen. So the slides in one trough of ultra pure water at 37°C to warm (See note ii). band(s)/staining that is competed by the antigen/peptide is specific. If more than one band disappears by peptide/antigen competition then those 3. Remove the other trough and into this dissolve the calcium chloride and bands have the antigenic determinants and could be considered either chymotrypsin using a magnetic stirrer (See note iii). Once dissolved, pH to fragments of the large antigen or multimer. 7.8 using the 0.5% sodium hydroxide and hydrochloric acid solutions. Return the trough to the water bath and allow this enzyme solution to re- 1. Determine the amount of antibody that is needed for 1 strip (e.g., 2 ml).

heat to 37°C (See note iv). Neuroscience protocols and tips For example, an antibody has given desired bands at 1:1000 dilution. So you will need 1 µl/ml antibody (2 µl antibody for 2 ml antibody solution). If 4. Transfer the warmed slides into the enzyme solution for a suggested 20 an antibody was used at 1:5000 dilution then you would only need 0.2 minutes (See note v) then remove the slides and place them into cold µl/ml (use 2 µl of 1:10 dilution for better accuracy). running tap water for 3 minutes (See note vi). 2. Take 2 µl antibody (or as needed) in 100 µl saline/PBS. Make 2 tubes. 5. Continue with immunohistochemical staining protocol. Add antigen/peptide solution (10-50 µg peptide or antigen added in 10-100 µl). Add same volume of saline/PBS (no peptide/antigen) to the other tube Notes labeled as “no peptide”. Mix gently. i. Use a sufficient volume of ultra pure water in order to cover the slides. Adjust the reagent quantities appropriately (the recipe above in the 3. Incubate both tubes at 37˚C for 1-2 hrs or 2-24 hrs at 4˚C. “Reagents” section is for 100 ml). 4. Centrifuge the tubes for 15 min at 4˚C in a microfuge (10-15000 rpm) to ii. Placing cold slides into the enzyme solution will lower the temperature of pellet any immune complexes. Carefully remove the supernatant. If no the solution, therefore reducing enzyme activity leading to the antigens visible pellet is seen then just leave ~ 5-10 µl at the bottom to avoid being under-retrieved. Once the slides have been de-waxed and re- disturbing invisible immune complexes. If you do not centrifuge the hydrated, drying out will cause non-specific antibody binding and therefore solution, it may give high background. high background staining. 5. After centrifugation, make up the volume of supernatant to 2 ml (or what iii. Chymotrypsin can be very allergenic. Use a face mask and extraction is necessary depending upon the initial antibody taken) with buffer (PBS- cabinet for weighing out. Tween with or without BSA or milk or any buffer that was used initially). Use both antibodies (with and without antigen/peptide) for Western blotting iv. Prepare the chymotrypsin solution as quickly as possible to avoid or immunolocalization. impairing the activity of the enzyme. Allow this solution to return to 37°C before introducing the slides. 6. Observe the bands/staining that disappears. v. Twenty minutes is only suggested as a starting point incubation time. Less than 20 minutes may leave the antigens under retrieved, leading to X. IHC-PARAFFIN PROTOCOLS (IHC-P) weak staining. More than 20 minutes may leave them over retrieved, leading to non-specific background staining and also increasing the A) Antigen retrieval chances of sections dissociating from the slides. A control experiment is Formalin fixed tissue requires an antigen retrieval step before recommended beforehand, where slides of the same tissue section are immunohistochemical staining can proceed. This is due to the formation of incubated in the enzyme solution for 10, 15, 20, 25, and 30 minutes before methylene bridges during fixation, which cross link proteins and therefore being immunohistochemically stained to evaluate optimum antigen mask antigenic sites. The two methods of antigen retrieval are enzymatic retrieval time for the particular antibody being used. and heat mediated. Both serve to break the methylene bridges and so expose the antigenic sites in order to allow the antibodies to bind. Some vi. Tap water stops the enzymatic process by washing action. antigens prefer enzymatic to heat mediated antigen retrieval and vice versa. Enzymatic tends to be a much gentler process than heated ii) Pressure Cooker Method mediated, so is best suited to more friable tissues. However, enzymatic Tissue sections are best mounted on APES (amino-propyl-tri-ethoxy- tends to take longer and is more technically demanding. silane) coated slides. Slides should be placed in a metal rack for this If no antigen retrieval step is stated on the antibody data sheet, start off by procedure. trying the heat mediated pressure cooker method.

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Materials Materials • Domestic stainless steel pressure cooker • Domestic (850W) or scientific microwave • Hot plate • pH meter • pH meter • Magnetic stirrer • Magnetic stirrer • 1 L beaker or conical flask • 2 L beaker or conical flask • Microwaveable vessel, either in-built or to hold approximately 400-500 ml

Reagents Reagents • Tri-Sodium Citrate 5.88 g • Tri-Sodium Citrate 2.94 g • 0.2 M Hydrochloric acid solution 44 ml • 0.2 M Hydrochloric acid solution 22 ml • Ultra-pure water 1956 ml • Ultra-pure water 978 ml • 0.5% sodium hydroxide solution • 0.5% sodium hydroxide solution • 0.5% hydrochloric acid solution • 0.5% hydrochloric acid solution • Xylene • Xylene • Industrial methylated spirits (IMS) or methanol • Industrial methylated spirits (IMS) or methanol

Method Method 1. Add the tri-sodium citrate, hydrochloric acid and ultra-pure water 1. De-wax and re-hydrate the paraffin sections by placing them in 3 together in a 2 L beaker/conical flask. Use a magnetic stirrer to ensure that changes of xylene for 3 minutes each, followed by 3 changes of IMS or all reagents are properly dissolved. methanol for 3 minutes each, followed by cold running tap water. Keep them in the tap water until the microwave antigen retrieval solution has 2. Adjust to pH 6.0 with 0.5% sodium hydroxide and hydrochloric acid been prepared. At no time from this point onwards should the slides be solutions. Add this solution to the pressure cooker. Place the pressure allowed to dry out! (See note i) cooker on the hotplate and turn it on full power. Do not secure the lid of the pressure cooker at this point, simply rest it on top. 2. Add the tri-sodium citrate, hydrochloric acid and ultra-pure water together in a 1 L beaker/conical flask. Use a magnetic stirrer to ensure that 3. While waiting for the pressure cooker to come to a boil, de-wax and re- all reagents are properly dissolved. Adjust to pH 6.0 with 0.5% sodium hydrate the paraffin sections by placing them in 3 changes of xylene for 3 hydroxide and hydrochloric acid solutions. Add this solution to the minutes each, followed by 3 changes of IMS or methanol for 3 minutes microwaveable vessel (See note ii). each, followed by cold running tap water. Keep them in the tap water until the pressure cooker comes to a boil. At no time from this point onwards 3. Remove the slides from the tap water and place them in the should the slides be allowed to dry out! (See note i.) microwaveable vessel. Place the vessel inside the microwave. If domestic, set to full power and wait until the solution comes to the boil. Boil for 15 4. Once boiling, transfer the slides from the tap water to the pressure minutes from this point. If scientific, program so that antigens are retrieved cooker. USE CARE WITH HOT SOLUTION - USE FORCEPS! Secure the for 15 minutes once the temperature has reached 98°C. (See note iii). pressure cooker lid as in the manufacturer’s instructions. 4. When 15 minutes has elapsed, remove the vessel and run cold tap 5. As soon as the cooker has reached full pressure (see the manufacturers water into it for 10 minutes. USE CARE WITH HOT SOLUTION! (See note iv). instructions), time 3 minutes. (See note ii). 5. Continue with immunohistochemical staining protocol. 6. When 3 minutes has elapsed, turn off the hotplate and place the pressure cooker in an empty sink. Activate the pressure release valve (see Notes the manufacturer’s instructions) and run cold water over the cooker. Once i. Once the slides have been de-waxed and re-hydrated, drying out will de-pressurized, open the lid and run cold water into the cooker for 10 cause non-specific antibody binding and therefore high background staining. minutes. CARE WITH HOT SOLUTION! (See note iii). ii. Use a sufficient volume of antigen retrieval solution in order to cover the 7. Continue with immunohistochemical staining protocol. slides. This should be by at least a few cm if using a non-sealed vessel to allow for evaporation during the boil. Notes i. Once the slides have been de-waxed and re-hydrated, drying out will iii. 15 minutes is only a suggested antigen retrieval time. Less than 15 cause non-specific antibody binding and therefore high background minutes may leave the antigens under retrieved, leading to weak staining. staining. More than 15 minutes may leave them over retrieved, leading to non-

Neuroscience protocols and tips specific background staining and also increasing the chances of sections ii. 3 minutes is only suggested as a starting point antigen retrieval time. dissociating from the slides. A control experiment is recommended Less than 3 minutes may leave the antigens under retrieved, leading to beforehand, where slides of the same tissue section are retrieved for 5, weak staining. More than 3 minutes may leave them over retrieved, 10, 15, 20, 25 and 30 minutes before being immunohistochemically leading to non-specific background staining and also increasing the stained to evaluate optimum antigen retrieval time for the particular chances of sections dissociating from the slides. A control experiment is antibody being used. recommended beforehand, where slides of the same tissue section are retrieved for 1, 2, 3, 4 and 5 minutes before being immunohistochemically iv. This is to make the slides cool enough to handle and to allow the stained to evaluate optimum antigen retrieval time for the particular antigenic site to re-form after being exposed to such high temperature. antibody being used. B) ABC Immunohistochemical Staining - iii. This is to make the slides cool enough to handle and to allow the antigenic site to re-form after being exposed to such high temperature. Chromogenic detection General Guidelines Carry out all incubations in a humidified chamber so that the sections do iii) Microwave Method not dry out. At no stage during the protocol should drying of the sections The use of a domestic microwave is inadvisable. Hot and cold spots are be allowed to occur. Drying out will lead to non-specific binding and common, leading to uneven antigen retrieval. Antigen retrieval times are ultimately high background staining. A shallow, plastic “sandwich type” box usually longer, due to the absence of a pressurized environment, nearly with a sealed lid and wet tissue paper in the bottom will be adequate, just always leading to section dissociation. as long as the slides can lay flat so that the reagents don’t drain off! A scientific microwave is much more appropriate. Most brands have on- board pressurized vessels and can keep the temperature at a constant A better solution is to cut a plastic serological pipette into lengths to fit your 98°C to avoid section dissociation. The only drawback is the expense of incubation chamber. Glue them in pairs to the bottom of the chamber, with purchasing one! the 2 individual pipette tubes of each pair being placed about 4.0 cm apart. Tissue sections are best mounted on APES (amino-propyl-tri-ethoxy- This provides a level and raised surface for the slides to rest on away from silane) coated slides. Slides should be placed in a plastic rack for this the wet tissue paper. procedure.

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 76 Neuroscience resources: www.abcam.com/neuroscience Carry out all TBS rinses between reagents in an appropriate glass trough. If the secondary antibody did have an affinity for any of the endogenous immunoglobulins on the mouse tissue, it can no longer bind to them as Dilutions of the primary antibody, secondary antibody and the ABC they have already been bound to immunoglobulins in the goat serum. complex should be determined by the individual product’s data sheet or Immunoglobulins from the same species will not interact with each other from past experience. Adjust dilutions appropriately from the results and therefore the secondary can only bind to the primary antibody. obtained. The use of normal serum before the application of the primary also Adhere strictly to all incubation times in the protocol eliminates Fc receptor binding of both the primary and secondary antibody.

Note: The main body of this protocol focuses on an ABC-Enzymatic The BSA serves a similar purpose as the normal serum, but this works to method (3 step). However, this protocol can be easily adapted for an ABC- reduce non-specific binding caused by hydrophobic interactions. Fluorescent method and also a 2 step or 1 step enzymatic or fluorescent method. See the “Protocol Adaptations” section for details. Cases can occur where the primary antibody recognizes another epitope showing sufficient homology with its actual target, for example a non- Protocol related tissue component. This type of background is greatly reduced by Please refer to Notes section for the theory and the Reagents section for the careful screening of monoclonal antibodies and the absorption of the recipes: polyclonal antibodies. General non-specific binding of Abcam primary antibodies is now rare for similar reasons. Perform antigen retrieval before commencing with the following protocols! Monoclonal antibodies tend to show less cross-reactivity with closely homologous epitopes than polyclonals, due to each monoclonal antibody DAY 1: molecule recognizing exactly the same epitope. 1. Rinse 2 x 5min TBS 2. 1.6% H2O2 in TBS for 30 min (HRP only) (See note i) iv. Simply remove the excess serum so not to further dilute the primary 3. Rinse 2 x 5min TBS 0.025% Triton (See note ii) antibody. 4. Block in 10% NS with 1% BSA in TBS for 2 hours at room temperature (See note iii) v. The primary antibody will target the epitope it is raised against. For example, an amino acid sequence unique to the CD4 molecule. 5. Drain slides for a few seconds (do NOT rinse) and wipe round sections (See note iv) Make sure that the primary antibody is raised in a different species to the 6. Apply primary antibody made up in TBS with 1% BSA. (See note v) tissue being stained. If, for example, you had mouse tissue and your 7. Incubate overnight at 4˚C, preferably on an orbital shaker primary antibody was raised in a mouse, the application of an anti-mouse (GENTLY!) (See note vi) IgG secondary antibody would bind to all the endogenous IgG in the mouse tissue. High non-specific background would occur. As discussed earlier, pre-incubation of the tissue with normal mouse serum would have DAY2:

no effect due to immunoglobulins from the same species not interacting Neuroscience protocols and tips 8. Rinse 2 x 5min TBS 0.025% Triton with each other 9. Apply secondary biotinylated antibody made up in TBS with 1% BSA for 2 hours at room temperature (AT THIS POINT MAKE UP THE vi. Overnight incubation allows antibodies of lower titer or affinity to be ABC COMPLEX IN TBS) (See note vii and viii) used by simply allowing more time for the antibodies to bind. Also, 10. Rinse 2 x 5min TBS whatever the antibody’s titer or affinity for its target, once the tissue has 11. Apply ABC complex in TBS for 30 min at room temperature (See note ix) reached saturation point no more binding can take place. Overnight 12. Rinse 2 x 5min TBS incubation assures that this occurs. 13. Develop with chromogen for 10 min at room temperature (See note x) 14. Rinse in running tap water for 5 min. The lower incubation temperature and gentle agitation from an orbital 15. Counterstain (If required) shaker is believed to help reduce background staining by increasing 16. Dehydrate, clear and mount (See note xi and xii) reaction times.

Notes vii. The secondary antibody recognizes the immunoglobulin species and i. H2O2 suppresses endogenous peroxidase activity and therefore reduces subtype of the primary antibody. background staining. Using a low concentration for 10 minutes adequately blocks endogenous peroxidase activity without having a detrimental effect In this example, a biotinylated Goat anti-Rabbit IgG antibody is being used on tissue epitopes. to bind to the primary Rabbit IgG anti-Mouse CD4 antibody. If using AP, then omit this step and step 1). See note x for further details. The secondary antibody is biotinylated, meaning that it has been conjugated with biotin. ii. The use of 0.025% Triton in the TBS helps to reduce surface tension, allowing reagents to cover the whole tissue section with ease. It is also When applied, the ABC complex will bind to this secondary biotinylated believed to dissolve Fc receptors, therefore reducing non-specific binding. antibody. Abcam recommends TBS to give a cleaner background than PBS viii. After the addition of the secondary antibody, make up the ABC iii. The secondary antibody may cross react with endogenous complex and leave to stand for a minimum of 30 minutes. This is the immunoglobulins in the tissue. This is avoided by pre-treating the tissue length of time that the complex takes to form. with normal serum from the species in which the secondary was raised. ix. The ABC complex consists of biotin-HRP/AP conjugate bound to avidin. For example, when detecting CD4 on mouse tissue and the secondary is raised in a goat: Avidin is a protein found in chicken egg white and has similar properties to streptavidin, a protein found in Streptomyces avidinii. Both Avidin and Firstly, block in normal goat serum. This will bind to any mouse streptavidin have a high affinity for biotin, a co-factor in enzymes involved immunoglobulins that show cross reactivity with goat immunoglobulins. in carboxylation reactions. On addition of the primary antibody, for example a Rabbit IgG anti-Mouse Both avidin and streptavidin can be used in an IHC application, but CD4, this antibody will bind to its target epitope on the CD4 molecule. streptavidin tends to be favored since it shows greater sensitivity. Streptavidin also produces less non-specific background staining, due to it On addition of the secondary antibody, for example a biotinylated Goat being non-glycosylated (unlike avidin), so shows no interaction with lectins anti-Rabbit IgG, this antibody will now only bind to the primary Rabbit IgG or other carbohydrate binding proteins. anti-Mouse CD4 antibody.

Customer Service Tel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM) 77 The complex is formed through avidin having 4 binding sites for biotin. Due D) IHC troubleshooting tips to steric hindrance, only 3 or fewer biotin molecules can actually bind. The ABC complex binds to the biotinylated secondary antibody, which is bound No staining to the primary antibody, which is in turn bound to the target on the tissue section. The primary antibody and the secondary antibody are not compatible. Use secondary antibody that was raised against the species in which the Due to the lack of BSA, the surface tension of the ABC solution is greater primary was raised (e.g primary is raised in rabbit, use anti-rabbit secondary). than the other reagents and will need spreading out over the tissue. A 200 µl pipette tip is good for this. Not enough primary antibody is bound to the protein of interest. x. Develop the colored product of the enzyme with the appropriate Use less dilute antibody, Incubate longer (e.g. overnight) at 4˚C. chromogen. The choice depends on which enzyme label you are using, the colored end product you prefer and whether you are using aqueous or organic mounting media (See note xi) for further details): The antibody may not be suitable for IHC procedures which reveal the protein in its native (3D form).

HRP AP Test the antibody in a native (non-denatured) WB to make sure it is not damaged. AEC (red) Fast Red (pink) The primary/secondary antibody/amplification kit may have lost its DAB (brown) INT (yellow/brown) activity due to improper storage, improper dilution or extensive NBT (purple/brown) freezing/thawing.

New Fuchsin (red) Run positive controls to ensure that the primary/secondary antibody is working properly. TNBT (purple) The protein is not present in the tissue of interest. Vega Red (pink) Run a positive control recommended by the supplier of the antibody. xi. Don’t forget that DAB is a suspected carcinogen. Wear the appropriate protective clothing. Deactivate it with chloros in a sealed container The protein of interest is not abundantly present in the tissue. overnight (it produces noxious fumes when chloros is added) and dispose of it according to laboratory guidelines. Use an amplification step to maximize the signal.

If using AP, add 0.24 mg/ml Levamisole (Sigma L9756) to the chromogen The secondary antibody was not stored in the dark. solution. Levamisole suppresses endogenous phosphatase activity and therefore reduces background staining. Always prevent the secondary antibody from exposure to light.

xii. If using AEC, Fast Red, INT or any other aqueous chromogen then Deparafinization may be insufficient. don’t forget that they are alcohol soluble. Use a suitable aqueous mounting media. Don’t dehydrate and clear! Deparaffinize sections longer, change the xylene.

xiii. Dehydrate and clear DAB, New Fuchsin, Vega Red, NBT, TNBT or any Fixation procedures (using formalin and paraformaldehyde fixatives) other organic chromogen developed sections by sending them through 3 may be modifying the epitope the antibody recognizes. changes of methanol (or IMS) for 3 minutes each followed by 3 changes of xylene for 3 minutes each. Mount sections in a suitable organic mounting Use antigen retrieval methods to unmask the epitope, fix for less time. media. Sections mounted in organic mounting media have a better refractive index than those mounted in aqueous mounting media. This The protein is located in the nucleus and the antibody (Nuclear protein) means that the image seen down the microscope will be sharper and cannot penetrate the nucleus. clearer if organic mounting media is used. Add a permeabilizing agent to the blocking buffer and antibody dilution buffer. Conclusion You should now see a colored product localized at the site of antibody The PBS buffer is contaminated with bacteria that damage the Neuroscience protocols and tips binding. phosphate groups on the protein of interest.

This corresponds to the location of your target. Add 0.01% azide in the PBS antibody storage buffer or use fresh sterile PBS. C) ABC Immunohistochemical staining - fluorescent detection High background - Possible reasons and solutions

Perform antigen retrieval before commencing with the Blocking of non specific binding might be absent or insufficient. following protocols! Increase the blocking incubation period and consider changing blocking agent. Abcam recommends 10% normal serum 1hr for sections or 1-5% DAY 1: BSA for 30 min for cells in culture. 1. Rinse 3 x 5min PBS 2. Rinse 2 x 5min PBS 0.025% Triton The primary antibody concentration may be too high. 3. Apply primary antibody - enzyme conjugate made up in PBS with 1% BSA Titrate the antibody to the optimal concentration, incubate for longer but in 4. Incubate overnight at 4˚C, preferably on an orbital shaker more dilute antibody (a slow but targeted binding is best). DAY 2: Incubation temperature may be too high. 6. Rinse 3 x 5min PBS 7. Incubate with secondary anitbody which is fluorescently labelled Incubate sections or cells at 4°C. 8. Rinse 3 x 5min PBS 9. Mount The secondary antibody may be binding non-specifically (damaged).

Run a secondary control without primary antibody.

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 78 Neuroscience resources: www.abcam.com/neuroscience High background XI. IMMUNOHISTOCHEMISTRY (IHC-FR) - FROZEN Tissue not washed enough, fixative still present. SECTIONS PROTOCOL

Wash extensively in PBS between all steps. Frozen sections: Once mounted on APES coated slides, frozen sections they are best kept at -80˚C until needed. Endogenous peroxidases are active. 1. When required, leave to warm at room temperature for 5 min. Use enzyme inhibitors i.e. Levamisol (2 mM) for alkaline phosphatase or 2 2 H O (0.3% v/v) for peroxidase. (See IHC protocol). 2. Pre-cool the fixative (acetone, methanol or ethanol) (Abcam recommends starting with acetone) at -20˚C for 30 min. Fixation procedures (using formalin and paraformaldehyde fixatives) are too strong and modified the epitope the antibody recognizes. 3. Fix with the pre-cooled fixative for 5 -10 min, at room temperature Change antigen retrieval method, decrease the incubation time with the 4. Rinse 3-4 X in PBS. antigen unmasking solution. 5. Continue with the immunohistochemical staining protocol. The absence Too much amplification (Amplification technique). of formalin eliminates the need for an antigen retrieval step. However, if frozen tissue or cytological specimens have been fixed in formalin, Reduce amplification incubation time and dilute the amplification kit. antigen retrieval can be attempted although the friable nature of the specimens may compromise the success. Too much substrate was applied (enzymatic detection). See section X, A) and B) for detailed protocols for chromogenic and Reduce substrate incubation time. fluorescent detection. The chromogen reacts with the PBS present in the cells/tissue (enzymatic detection). XII. IMMUNOCYTOCHEMISTRY (ICC) PROTOCOL Use Tris buffer to wash sections prior to incubating with the substrate, then wash sections/cells in Tris buffer. i Coat coverslips with polyethylineimine or poly-L-lysine for 1 hr at room temperature. Pemeabilization has damaged the membrane and removed the ii Rinse coverslips well with sterile H2O (3 times 5 min each). membrane protein (membrane protein). iii Allow coverslips to dry completely and sterilize them under UV light for at least 4 hrs. Remove permeabilizing agent from your buffers. iv Grow cells on glass coverslips or prepare cytospin or smear preparation. v Rinse briefly in phosphate-buffered saline (PBS). Neuroscience protocols and tips Non-specific staining Fixation: 1. Fix the samples either in ice-cold methanol, acetone (1-10 min) or in 3- Primary/secondary antibody concentration may be too high. 4% paraformaldehyde in PBS pH 7.4 for 15 min at room temperature. Try decreasing the antibody concentration and/or the incubation period. Compare signal intensity against cells that do not express the target. 2. Wash the samples twice with ice cold PBS.

Endogenous peroxidases are active. Permeabilization: If the target protein is expressed intracellularly, it is very important to Use enzyme inhibitors i.e. Levamisol (2 mM) for alkaline phosphatase or permeabilize the cells. Note: acetone fixed samples do not require H2O2 (0.3% v/v) for peroxidase. (See IHC protocol). permeabilization.

3. Incubate the samples for 10 min with PBS containing 0.25% Triton X- The primary antibody is raised against the same species as the 100 (or 100 µM digitonin or 0.5% saponin). Triton X-100 is the most tissue stained (e.g Mouse primary antibody tested on mouse tissue). popular detergent for improving the penetration of the antibody. However, it When the secondary antibody is applied it binds to all the tissue as it is not appropriate for the use of membrane-associated antigens since it is raised against that species. destroys membranes. Use a primary antibody raised against a different species than your tissue. 4. Wash cells in PBS three times for 5 min. The sections/cells have dried out. Blocking and Incubation: 5. Incubate cells with 1% BSA in PBST for 30 min to block unspecific Keep sections/cells at high humidity and do not let them dry out. binding of the antibodies (alternative blocking solutions are 1% gelatin or 10% serum from the species that the secondary antibody was raised in).

6. Incubate cells in the diluted antibody in 1% BSA in PBST in a humidified chamber for 1 hr at room temperature or overnight at 4˚C.

7. Decant the solution and wash the cells three times in PBS, 5 min each wash. Brighten 8. Incubate cells with the secondary antibody in 1% BSA for 1 hr at room temperature in dark. up your 9. Decant the secondary antibody solution and wash three times with PBS Neurons! for 5 min each in dark.

Counter staining: 10. Incubate cells on 0.1-1 µg/ml Hoechst or DAPI (DNA stain) for 1 min.

11. Rinse with PBS.

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Mounting: Red-conjugated against rabbit and FITC-conjugated against mouse) in 12. Mount coverslip with a drop of mounting medium. 1% BSA for 1 hr at room temperature in dark.

13. Seal coverslip with nail polish to prevent drying and movement under 9. Decant the mixture of the secondary antibody solution and wash three microscope. times with PBS for 5 min each in dark.

14. Store in dark at -20 or 4˚C. Counter staining: 10. Incubate cells on 0.1-1 µg/ml Hoechst or DAPI (DNA stain) for 1 min.

XIII. DOUBLE IMMUNOFLUORESCENCE- 11. Rinse with PBS. SIMULTANEOUS (IHC-P, IHC-FR, ICC) PROTOCOL Mounting: 12. Mount coverslip with a drop of mounting medium. In order to be able to examine the co-distribution of two (or more) different antigens in the same sample, a double immunofluorescence procedure 13. Seal coverslip with nail polish to prevent drying and movement under can be carried out. Primary antibodies raised in different species can be microscope. used either in parallel (in a mixture) or in a sequential way. 14. Store in dark at -20˚C or 4˚C. Preparation of slides and samples:

A. Cell lines, cytology smears, cytospin preparations i. Coat coverslips with polyethylineimine or poly-L-lysine for 1 hr at room XIV. DOUBLE IMMUNOFLUORESCENCE- temperature. SEQUENTIAL PROTOCOL ii. Rinse coverslips well with sterile H2O (3 times 5 min each). iii. Allow coverslips to dry completely and sterilize them under UV light Fixation and permeabilization, are the same as for part XIII. for at least 4 hrs. iv. Grow cells on glass coverslips or prepare cytospin or smear preparation. Blocking and Sequential Incubation: v. Rinse briefly in phosphate-buffered saline (PBS). 1. First blocking step: incubate cells with the first serum (10% serum B. Frozen (cryostat) sections from the species that the secondary antibody was raised in) for 30 min to i. Snap frozen fresh tissue in liquid nitrogen or isopentane pre-cooled in block unspecific binding of the antibodies (alternative blocking solutions liquid nitrogen. Store frozen blocks at -80˚C. are 1% gelatin or 1% BSA) at room temperature. ii. Cut 4-8 µm thick cryostat sections and mount on superfrost or gelatin coated slides. You can store slides at -80˚C until needed. 2. Incubate cells with the first primary antibody in 1% BSA or 1% serum iii. Before IF staining, warm up slides at room temperature for 30 minutes. in PBST in a humidified chamber for 1 hr at room temperature or overnight at 4˚C depending on the concentration of the antibody and the accessibility C. Paraffin-embedded sections of the antigen. i. Deparaffinize sections in xylene 2x5 min. ii. Hydrate with 100% ethanol 2x3 min. 3. Decant the first primary antibody solution and wash the cells three times iii. Hydrate with 95% ethanol 1 min. in PBS, 5 min each wash. iv. Rinse in distilled water and then follow procedure for fixation and antigen retrieval as required (please see IHC protocol for formalin- 4. Incubate cells with first secondary antibody (labelled with fixed paraffin-embedded tissue sections for further details). Fluorochrome-1) in 1% BSA in PBST for 1 hr at room temperature in dark.

Fixation: 5. Decant the first secondary antibody solution and wash three times with 1. Fix the samples either in ice-cold methanol, acetone (1-10 min) or in 3- PBS for 5 min each in dark. 4% paraformaldehyde in PBS pH 7.4 for 15 min at room temperature. 6. Second blocking step: incubate cells with the second serum (10% 2. Wash the samples twice with ice cold PBS. serum from the species that the secondary antibody was raised in) for 30 min to block unspecific binding of the antibodies (alternative blocking Permeabilization: solutions are 1% gelatin or 1% BSA) at room temperature in the dark. If the target protein is expressed intracellularly, it is very important to permeabilize the cells. Note: acetone fixed samples do not require 7. Incubate cells with the second primary antibody in 1% BSA in PBST Neuroscience protocols and tips permeabilization. in a humidified chamber in the dark for 1 hr at room temperature or overnight at 4˚C depending on the concentration of the antibody and the 3. Incubate the samples for 10 min with PBS containing 0.25% Triton X- accessibility of the antigen. 100 (or 100 µM digitonin or 0.5% saponin). Triton X-100 is the most popular detergent for improving the penetration of the antibody. However, it 8. Decant the second primary antibody solution and wash the cells three is not appropriate for the use of membrane-associated antigens since it times in PBS, 5 min each wash in dark. destroys membranes. 9. Incubate cells with second secondary antibody (labelled with 4. Wash cells in PBS three times for 5 min. Fluorochrome-2) in 1% BSA for 1 hr at room temperature in dark.

Blocking and Simultaneous Incubation: 10. Decant the second secondary antibody solution and wash three times 5. Incubate cells with 1% BSA in PBST for 30 min to block unspecific with PBS for 5 min each in dark. binding of the antibodies (alternative blocking solutions are 1% gelatin or 10% serum from the species that the secondary antibody was raised in). Counter staining: 1. Incubate cells on 0.1-1 µg/ml Hoechst or DAPI (DNA stain) for 1 min in dark. 6. Incubate cells in the mixture of two primary antibodies (e.g. rabbit against human target-1 and mouse against human target-2, if the targets 2. Rinse with PBS in dark. are human proteins) in 1% BSA in PBST in a humidified chamber for 1 hr at room temperature or overnight at 4oC. Mounting: Mount coverslip with a drop of mounting medium. 7. Decant the mixture solution and wash the cells three times in PBS, 5 min each wash. 1. Seal coverslip with nail polish to prevent drying and movement under microscope. 8. Incubate cells with the mixture of two secondary antibodies which are raised in different species (with two different fluorochromes, i.e. Texas 2. Store in dark -20˚C or 4˚C.

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 80 Neuroscience resources: www.abcam.com/neuroscience Fluorophores: 2. Add 0.1-10 µg/ml of the primary antibody. Dilutions, if necessary, should Absorbance Emission Visible color be made in 3% BSA/PBS. Dye Wavelength Wavelength 3. Incubate for at least 30 min at room temperature. Hydroxycoumarin 325 386 blue methoxycoumarin 360 410 blue 4. Wash the cells 3X by centrifugation at 400 g for 5 min and resuspend them in ice cold PBS. Alexa fluor 345 442 blue aminocoumarin 350 445 blue 5. Dilute the fluorochrome-labeled secondary antibody in 3% BSA/PBS at Cy2 490 510 green (dark) the optimal dilution (according to the manufacturer’s instructions) and then resuspend the cells in this solution. FAM 495 516 green (dark) Alexa fluor 488 494 517 green (light) 6. Incubate for at least 20 minutes at room temperature. This incubation Fluorescein FITC 495 518 green (light) must be done in the dark. Alexa fluor 430 430 545 green (light) 7. Wash the cells 3x as in step 4. Alexa fluor 532 530 555 green (light) HEX 535 556 green (light) 8. The cells are now ready for analysis by flow cytometry. Keep the cells on ice until your scheduled time for analysis. Cy3 550 570 yellow TRITC 547 572 yellow Remember that detection of intracellular antigens requires a cell Alexa fluor 546 556 573 yellow permeabilization step prior to staining. Alexa fluor 555 556 573 yellow R-phycoerythrin (PE) 480;565 578 yellow Rhodamine Red-X 560 580 orange XVI. INDIRECT FLOW CYTOMETRY (FACS) PROTOCOL

Tamara 565 580 red Procedure: Cy3.5 581 581 596 red Indirect labeling requires two incubation steps; one with a primary antibody Rox 575 602 red then the next one with a compatible secondary antibody. The secondary (and not the primary) antibodies have the fluorescent dye (FITC, PE, Cy5, Alexa fluor 568 578 603 red etc.) attached. Please note that this is a general protocol and you may Red 613 480;565 613 red need to adjust it for your applications. Neuroscience protocols and tips Texas Red 615 615 red 1. Harvest, wash the cells and determine the total cell number. Use Alexa fluor 594 590 617 red polystyrene round-bottom 12x75 mm Falcon tubes for Flow Alexa fluor 633 621 639 red cytometry/FACS analysis. It is always useful to check the viability of the cells which should be around 95% not less than 90%. Allophycocyanin 650 660 red Alexa fluor 633 650 668 red 2. Resuspend the cells approximately 1-5x107 cells/ml in ice cold 3% BSA/PBS. Cy5 650 670 red 3. Add 1 ml of cell suspension to each Falcon tube. Alexa fluor 660 663 690 red Cy5.5 675 694 red 4. Add 0.1-10 µg/ml of the primary antibody. Dilutions, if necessary, should TruRed 490;675 695 red be made in 3% BSA/PBS. Alexa fluor 680 679 702 red 5. Incubate for at least 30 min at room temperature. Cy7 743 770 red 6. Wash the cells 3-times by centrifugation at 400 g for 5 min and Nucleic acid probes: resuspend them in ice cold PBS. You may need to adjust the conditions of the centrifugation (the force and the time) for the cell types used. Absorbance Emission Visible color Dye Wavelength Wavelength 7. Dilute the fluorochrome-labeled secondary antibody in 3% BSA/PBS at DAPI 345 455 blue the optimal dilution (according to the manufacturer’s instructions) and then Hoechst 33258 345 478 blue resuspend the cells in this solution.

SYTOX blue 431 480 blue 8. Incubate for at least 20 minutes at room temperature. This incubation Hoechst 33342 343 483 blue must be done in the dark. YOYO-1 509 509 green 9. Wash the cells 3-times by centrifugation at 400 g for 5 min and SYTOX green 504 533 green resuspend them in ice cold PBS. TOTO 1, TO-PRO-1 509 533 green SYTOX orange 547 570 yellow The cells are now ready for analysis by flow cytometry. Keep the cells on ice until your scheduled time for analysis. If you need to wait longer than Chromomycin A3 445 575 yellow for an hour, you may need to fix the cells which can preserve them for at Mithramycin 445 575 yellow least several days. (This will stabilize the light scatter and inactivate most Propidium iodide 536 617 red biohazardous agents) Ethidium bromide 493 620 red 10. After the last step, centrifuge the cells and remove the liquid.

11. Add 0.5 to 1.0 ml of cold 0.5% paraformaldehyde solution and vortex XV. DIRECT FLOW CYTOMETRY (FACS) PROTOCOL immediately. 12. Store the cell suspension at 4°C in the dark. General Procedure: 1. Harvest, wash the cells and adjust cell suspension to a concentration of 1-5x106 cells/ml in ice cold 3% BSA/PBS. Use polystyrene round-bottom Remember that detection of intracellular antigens 12x75 mm Falcon tubes. requires a cell permeabilization step prior to staining (use 0.5% saponin or acetone or Leucoperm).

Customer Service Tel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM) 81 XVII. DIRECT ELISA PROTOCOL 3. Wash the plate four times with PBS.

Buffers and Reagents: 4. Add 100 µl of conjugated secondary antibody, diluted at the optimal Bicarbonate/carbonate coating buffer (100 mM) - antigen or antibody concentration (according to the manufacturer) in blocking buffer immediately before use. should be diluted in coating buffer to immobilize them to the wells: 3.03 g Na2CO3, 6.0 g NaHCO3 (1000 ml distilled water) pH 9.6, PBS: 1.16 g 5. Cover the plate with an adhesive plastic and incubate for 1-2 h at room Na2HPO4, 0.1 g KCl, 0.1 g K3PO4, 4.0 g NaCl (500 ml distilled water) pH 7.4, temperature.

Blocking solution: commonly used blocking agents are BSA, nonfat dry 6. Wash the plate four times with PBS. milk, casein, gelatin. PBS containing 1% Bovine Serum Albumin (BSA), Detection Wash solution: usually PBS or Tris-buffered saline (pH 7.4) with detergent Although many different types of enzymes have been used for detection, such as 0.05% (v/v) Tween20 (TBST) horse radish peroxidase (HRP) and alkaline phosphatase (ALP) are the two widely used enzymes employed in ELISA assay. It is important to Antibody solution: primary and secondary antibody should be diluted in 1x consider the fact that some biological materials have high levels of blocking solution to prevent nonspecific bindings endogenous enzyme activity (such as high ALP in alveolar cells, high peroxidase in red blood cells) and this may result in non-specific signal. If Coating antigen to microplate necessary, perform an additional blocking treatment with Levamisol (for 1. Dilute the antigen to a final concentration of 20 µg/ml in PBS. Coat the ALP) or with 0.3% solution of H2O2 in methanol (for peroxidase). wells of a PVC microtiter plate with the antigen by pipeting 50 µl of the antigen dilution per well. ALP substrate For most applications pNPP (p-Nitrophenyl-phosphate) is the most widely 2. Cover the plate with an adhesive plastic and incubate for 2 h at room used substrate.The yellow color of nitrophenol can be measured at 405 temperature. nm after 15-30 min incubation at room temperature. (This reaction can be stopped by adding equal volume of 0.75 M NaOH). 3. Remove the coating solution and wash the plate twice by filling the wells with 200 µl PBS. The solutions or washes are removed by flicking the plate HRP chromogens over a sink. The remaining drops are removed by patting the plate on a paper The substrate for HRP is hydrogen peroxide. Cleavage of hydrogen towel. peroxide is coupled to oxidation of a hydrogen donor which changes colour during reaction. Blocking 4. Block the remaining protein-binding sites in the coated wells by adding TMB (3,3’,5,5’-tetramethylbenzidine) add TMB solution to each well, 200 µl blocking buffer, 5% non fat dry milk/PBS, per well. Alternative incubate for 15-30 min, add equal volume of stopping solution (2 M H2SO4) blocking reagents include BlockACE or BSA. and read the optical density at 450 nm.

5. Cover the plate with an adhesive plastic and incubate for at least 2 h at OPD (o-phenylenediamine dihydrochloride. The end product is measured room temperature or, if more convenient, overnight at 4°C. at 492 nm. Be aware that the substrate is light sensitive so keep and store it in the dark. 6. Wash the plate twice with PBS. ABTS (2,2’-azino-di-[3-ethyl-benzothiazoline-6 sulfonic acid] diammonium Incubation with the Antibody salt. The end product is green and the optical density can be measured at 7. Add 100 µl of the antibody, diluted at the optimal concentration 416 nm. (according to the manufacturer’s instructions) in blocking buffer immediately before use. Note: some enzyme substrates are considered hazardous (potential carcinogens), therefore always handle with care and wear gloves. 8. Cover the plate with an adhesive plastic and incubate for 2 h at room temperature. 7. Dispense 100 µl (or 50 µl) of the substrate solution per well with a multichannel pipet or a multipipet. 9. Wash the plate four times with PBS. 8. After sufficient color development (if it is necessary) add 100 µl of stop Detection solution to the wells.

Neuroscience protocols and tips 10. Dispense 100 µl (or 50 µl) of the substrate solution per well with a multichannel pipet or a multipipet. 9. Read the absorbance (optical density) of each well with a plate reader.

11. After sufficient color development (if it is necessary) add 100 µl of stop solution to the wells. XIX. ELISA USING FLUORESCENT SUBSTRATE 12. Read the absorbance (optical density) of each well with a plate reader. PROTOCOL Note: some enzyme substrates are considered hazardous (potential carcinogens), therefore always handle with care and wear gloves. Day 1: 1. Coat with 100 µl/well of coating antibody diluted in filtered PBS. Incubate plate overnight at 4˚C, covered with plate sealer. XVIII. INDIRECT ELISA PROTOCOL Day 2: 2. Block plates with 200 µl/well of 4 g Block ACE powder, diluted in 100 ml Buffers and Reagents: (See section XVI Buffers and Reagents) of deionized water (USE a 1:4 dilution of this) for 3 hrs at RT covered with plate sealer. For accurate quantitative results, always compare signal of unknown samples against those of a standard curve. Standards (duplicates or 3. Wash plates: PBS-T (0.05% Tween20) 250 µl/well; 3x 30 seconds. triplicates) and blank must be run with each plate to ensure accuracy. 4. Load 100 µl of standards or samples ** freshly diluted in 10% BlockACE Incubation with Primary and Secondary antibody in PBS-T overnight at 4˚C, covered with plate sealer. 1. Add 100 µl of diluted primary antibody to each well. Day 3: 5. Incubate 100 µl/well of biotinylated reporter antibody diluted in PBS for 2. Cover the plate with an adhesive plastic and incubate for 2 h at room 2 hours at RT covered with plate sealer temperature.

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 82 Neuroscience resources: www.abcam.com/neuroscience 6. Incubate 100 µl/well of streptavidin alkaline phosphatase, 1:5000 24. Phenol:chloroform extract the samples and ethanol precipitate the dilution in PBS for 1 hour at RT, covered with plate sealer. DNA in presence of 10 µl glycogen (5 mg/ml). 25. Resuspend pellet in 100 µl H2O and store at -20°C or proceed with 7. Wash plates: TBS 250 µl/well; 3x 30 seconds. detection method (PCR, southern blot, etc). **

8. Amplify signal by adding 100 µl/well AttoPhos Fluorescent substrate * Chromatin can be snap frozen in liquid nitrogen after step 8 and stored system, for 5-10 min at RT. 36 mg of AttoPhos substrate should be mixed at -70˚C for up to 2 month. Avoid multiple freeze-thawing. with 60 ml of AttoPhos buffer 24 hours prior to use. Make sure well is ** The samples can be frozen and stored at -20˚C after these steps. clean-no contamination. Solutions 9. Signal measured on Fluorometer, (Victor2, Perkin Elmer); excitation: 440 nm; emission: 550 nm ChIP Lysis Buffer 50 ml Stock solution 50 mM HEPES-KOH 2.5 ml 1 M ** 140 mM NaCl 1.4 ml 5 M • Prepare standards ahead of time. 1 mM EDTA pH8 0.1 ml 0.5 M • On day of application to the plate, (day 2 in above procedures), 1% Triton X 2.5 ml 20% standards are freshly diluted in 10% BSA in PBS-T from 10 ng/ml to 500 pg/ml, for example. 0.1% Sodium Deoxycholate 0.5 ml 10% • After washing or aspirating flip plate over onto kim wipes on bench to Protease Inhibs. (added fresh each time) remove excess liquid. Dilution Buffer 50 ml Stock solution 1% Triton X-100 2.5 ml 20% XX. CHROMATIN IMMUNOPRECIPITATION (ChIP) 2 mM EDTA 0.2 ml 0.5 M PROTOCOL 150 mM NaCl 1.5 ml 5 M 20 mM Tris-HCl pH 8 1 ml 1 M Cross-linking and Cell Harvesting Protease Inhibs. (added fresh each time) 1. Start with 2 large dishes when confluent (1x107 _ 5x107 cells per dish). 2. Cross-link proteins to DNA by adding formaldehyde drop-wise directly to Wash Buffer 500 ml Stock solution media for a final concentration of 0.75% and rotate gently at room 0.1% SDS 2.5 ml 20% temperature (RT) for 10 min. 1% Triton X-100 25 ml 20% 3. Rinse cells 2 x with 10 ml cold PBS. 2 mM EDTA 2 ml 0.5 M 4. Scrape cells into 5 ml cold PBS and transfer into 50 ml tube. 150 mM NaCl 15 ml 5 M 5. Repeat with 3ml PBS. 20 mM Tris-HCl pH8 10 ml 1 M 6. Centrifuge for 5 min at 3,000 rpm, carefully aspirate off supernatant and 7 resuspend pellet in ChIP lysis buffer (750 µl per 1x10 cells). Final Wash Buffer 500 ml Stock solution Neuroscience protocols and tips Sonication 0.1% SDS 2.5ml 20% 7. Sonicate lysate to shear DNA to an average fragment size of 500 to 1% Triton X-100 25ml 20% 1000 bp. Follow the fragment size on a 1.5% agarose gel. 2 mM EDTA 2ml 0.5 M 8. Centrifuge for 30 secs at 13,000 rpm and transfer supernatant to new tube.* 500 mM NaCl 50ml 5 M 9. Remove 50 µl of each sonicated sample and add to 400 µI ChIP lysis buffer. 20 mM Tris-HCl pH8 10ml 1 M 10.This sample is the input. This is used for obtaining DNA concentration for subsequent IP’s (see below) and as control in the final PCRs. Elution Buffer 10 ml (make fresh each time) 1% SDS 0.5 ml 20% Determination of input DNA concentration 3 11. Add 5 µl of proteinase K (20 mg/ml) to each input sample in ChIP lysis 100 mM NaHCO 1 ml 1 M buffer and heat with shaking at 65°C for 4-5 hours (or overnight) to reverse Proteinase K (Roche) cross-linking.** Dissolve in H2O at 20 mg/ml, store at -20°C. 12. Phenol: chloroform extract the samples and ethanol precipitate the DNA in presence of 10 µl glycogen (5 mg/ml). Protein A/G beads with s/s Sonicated Salmon Sperm 13. Re-dissolve the pellet in 100 µl H2O. ** 1. Dissolve Salmon Sperm DNA (Sigma) at 5 mg/ml in 100 ml H2O (4x25 14. Transfer 5 µl of the sample in a tube containing 995 µl TE to give a ml). Rotate at 4°C overnight. 260 200-fold dilution and read the OD . The concentration of DNA in µg/ml is 2. Phenol:chloroform extract (1:1), spin for 2 min at RT at 5,000 rpm. 260 OD x 10,000. 3. Add 30 ml EtOH and 1.2 ml KAc (3 M) to each sample, precipitate, spin as above. Immunoprecipitation 15. Use 25 µg of chromatin per IP and dilute each sample 1:10 with 4. Wash with 70% EtOH, spin as above. dilution buffer. You will need one sample for the specific antibody, and one 5. Resuspend in H2O at 10 mg/ml (rotating at RT). sample for the beads-only control. 6. Sonicate, big probe, for 10-20 sec (check on agarose gel). 16. Add the primary antibody to all samples except the beads-only control. 7. Read OD260. Boil in screw-cap eppendorfs for 10-15 min then cool on ice. The amount of antibody to be added has to be determined empirically, 1-8 8. Mix equal volume of Protein A and Protein G beads. Wash 3x in PBS. µg per 25 µg of chromatin often works well. 9. Remove PBS and add s/s Salmon Sperm DNA (to 1.5 µg/20 µl beads). 17. Add 20 µl of protein A/G beads (pre-absorbed with sonicated s/s Add PBS back up to the original volume. Incubate for 30 min with rotation salmon sperm DNA at 1.5 µg/20 µl beads) to all samples and IP overnight at 4°C. with rotation at 4°C. 18. Centrifuge the protein A/G beads for 1 min at 3,500 rpm and remove the supernatant. XXI. BUFFERS AND STOCK SOLUTIONS 19. Wash beads 3 x with 1 ml wash buffer (centrifuge as above). 20. Wash beads 1 x with 1 ml final wash buffer (centrifuge as above). Cytoskeletal bound proteins Extract Buffer: 10 mM Tris, pH 7.4 Elution and reverse cross-link 100 mM NaCl 21. Elute DNA by adding 450 µl of elution buffer to the protein A/G beads 1 mM EDTA and rotate for 15 min at RT. 1 mM EGTA 22. Spin down and transfer the supernatant into fresh tube. ** 1 mM NaF 23. Reverse cross-links by adding 5 µl proteinase K (20 mg/ml) to the 20 mM Na4P2O7 eluates and heat with shaking at 65°C for 4-5 hours (or overnight ).** 2 mM Na3VO4

Customer Service Tel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM) 83 1% Triton X-100 Nuclear Fractionation Protocol Reagents 10% glycerol Buffer A – 10 mM HEPES, 1.5 mM MgCl2, 10 mM KCl, 0.5 mM DTT, 0.1% SDS 0.05% NP40 (or 0.05% Igepal or Tergitol) pH 7.9 0.5% deoxycholate To prepare 250 ml stock of buffer A – Soluble protein buffer: HEPES: 1M = 238.3 g/L, therefore 10 mM = 0.59 g/250 ml 20 mM Tris-HCl, pH 7.5 MgCl2: 1M = 203.3 g/L, therefore 1.5 mM = 0.076 g/250 ml 2+ 1 mM EGTA (Ca chelator) KCl: 1M = 74.5 g/L, therefore 10 mM = 0.187 g/250 ml DTT: 1M = 154.2 g/L, therefore 0.5 mM = 0.019 g/250 ml RIPA buffer (RadioImmunoPrecipitation Assay) buffer: NP40 = 0.05% RIPA buffer contains the ionic detergent sodium deoxycholate as an active constituent and is particularly used for nuclear membrane disruption for Buffer B – 5 mM HEPES, 1.5 mM MgCl2, 0.2 mM EDTA, 0.5 mM DTT, nuclear extracts. A RIPA buffer gives low background but can denature 26% glycerol (v/v), pH 7.9 kinases. It can also disrupt protein-protein interactions (and may therefore be problematic for immunoprecipitations/pull down assays). To prepare 250 ml stock of buffer B – HEPES: 1M = 238.3 g/L, therefore 5 mM = 0.295 g/250 ml 50mM Tris HCl pH 8 MgCl2: 1M = 203.3 g/L, therefore 1.5 mM = 0.076 g/250 ml 150 mM NaCl EDTA: 1M = 372.2 g/L, therefore 0.2 mM = 0.0186 g/250 ml 1% NP-40 DTT: 1M = 154.2 g/L, therefore 0.5 mM = 0.019 g/250 ml 0.5% sodium Deoxycholate 26% Glycerol (v/v) = 65 ml 0.1% SDS - 87.66 g/326 ml The 10% sodium deoxycholate stock solution (5 g into 50 ml) must be 4.6 M NaCl protected from light. TBS (Tris Buffered Saline) pH 7.6-7.8: The 100 mM EDTA stock solution is made with 1.86 g into 40 ml H2O and For 10 litres: 60.6 g TRIS HCl then add NaOH to dissolve and adjust pH to 7.4. Finally, adjust the total 13.9 g TRIS base volume to 50 ml). Store the buffer at 4˚C. 87.66 g NaCl Nonidet-P40 (NP-40) buffer: 10 litres Ultra pure water (H2O) 20 mM Tris HCl pH 8 137 mM NaCl 10% glycerol TBS 0.025% Triton X-100: 1% nonidet P-40 For 1 litre: 250 µl Triton X-100 2 mM EDTA 999.75 ml TBS pH 7.6-7.8 Sodium orthovanadate preparation: This needs to be done under the fume hood 1.6% H2O2 (Hydrogen Peroxide) in TBS: • Prepare a 100 mM solution in double distilled water • Set pH to 9.0 with HCl For 400 ml: 6.4 ml H2O2 (GPR = 30% w/w) • Boil until colorless • Cool to room temperature 393.6 ml TBS pH 7.6-7.8 • Set pH to 9.0 again • Boil again until colorless 10% NS (Normal Serum) with 1% BSA (Bovine Serum Albumin, • Repeat this cycle until the solution remains at pH 9.0 after boiling and Fraction 5) in TBS: cooling • Bring up to the initial volume with water For 1 ml: 100 µl NS • Store in aliquots at -20˚C 10 mg BSA Note: do not permit great changes in volume during boiling; put a loose lid 900 µl TBS pH 7.6-7.8 on the container to protect from evaporation. Discard if the samples turn yellow. Primary antibody made up in TBS with 1% BSA:

Neuroscience protocols and tips TBS 10x (concentrated TBS) (Example is of primary antibody used at a dilution of 1:10) 24.23 g Trizma HCl 100 µl Primary antibody 80.06 g NaCl For 0.1 ml: Mix in 800 ml ultra pure water. 10 mg BSA pH to 7.6 with pure HCl. 900 µl TBS pH 7.6-7.8 Top up to 1 L.

TBST Secondary biotinylated antibody made up in TBS with 1% BSA: For 1 L: 100 ml of TBS 10x + 900 ml ultra pure water + 1ml Tween20 (Example is of secondary biotinylated antibody used at a dilution of 1:200) For 1 ml: 5 µl Secondary biotinylated antibody Medium stripping buffer: Make fresh stripping buffer: 995 µl TBS pH 7.6-7.8 15 g glycine 1 g SDS ABC (Avidin-Biotin) complex in TBS: 10 ml Tween20 (Example is of ABC complex, each part used at a dilution of 1:100) Set the pH to 2.2 make up to 1 L with ultrapure water For 1 ml: 10 µl Streptavidin 10 µl HRP (or AP)-Biotin Harsh stripping buffer: to be done under the fumehood 980 µl TBS pH 7.6-7.8 For 100 ml: 20 ml SDS 10% Bicarbonate/carbonate coating buffer (100 mM): 3.03 g Na2CO3, 6.0 g 12.5 ml Tris HCl pH 6.8 0.5M NaHCO3 (1 L distilled water) pH 9.6, 67.5 ml ultra pure water PBS: 1.16 g Na2HPO4, 0.1 g KCl, 0.1 g K3PO4, 4 g NaCl (500 ml distilled Add 0.8ml ß-mercaptoethanol under the fumehood. water) pH 7.4

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 84 Neuroscience resources: www.abcam.com/neuroscience AD. LUMEN has - β γ ApoE, or A CYTOPLASM 2M), α β N T R 7 5 P A β A 4 4 C D β Protease NEP A TION Protease PLG I - A E E C D H R N β Alzheimer's Disease. A 2-macroglobulin ( secretase r b 4 α E B γ DEGRADA -secretase releases an amyloid T N C H γ O L P R Protease IDE β A Pen-2 ApoE -secretase (BACE1/2). Cleavage by BACE1 and then 1 2 β β A α α β Aph - 1 Aph - 1 Aph - 1 ), or by β 2M α Neuroscience signaling pathways EXPORT β A CLEARANCE / NCT s Disease , the major component of brain plaques characterising - secretase ’ cleavage activates γ β Protease A ApolipoproteinE (ApoE) in amyloid plaques, clearance by -secretase and the biological significance of these cleavages remain to be determined. γ or Mint - 3 P3 β Alzheimer β β A A A is also involved in an intracellular signaling cascade upon phosphorylation of tyrosine 688, the C- PS1 / PS2 β β β A A A ApoE APP . Ubiquilin β Mint - 2 A β Aph-1 and Pen-2) generates A Fe65 Mint - 1 CLAC-P OLIGOMERIZATION I A C D β T Y P Y E C N N fect nuclear transcription. Interactions of proteins such as presenilin (PS), with molecules ubiquilin, may contribute to A APP - 49 - 40 - 42 P T P B -secretase (ADAM10/17), precluding the formation of amyloid beta (A α Shc Grb2 BACE1 BACE2 ADAM10 ADAM17 SH2 sos Ras can be processed by - secretase - secretase - secretase Raf MAPK echnical help - Neuroscience signaling pathway diagrams β α γ terminus can be stabilized and linked to other proteins by interacting adaptor such as the Mints or Fe65 cleavage intracellular domain (AICD) that may af There are many other type-I integral membrane proteins that cleaved by Amyloid Precursor Protein (APP) processing in APP many fates: oligomerization and association with proteins such as CLAC-P low density lipoprotein receptor-related protein (LRP), or degradation by proteases such as insulin degrading enzyme (IDE), neprilysin (NEP) plasminogen (PLG). Alterations in any pathway can lead to increased levels of secretase (containing presenilin-1/2 (PS), Nicastrin, T

Customer Service Tel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM) 85 Technical help - Neuroscience signaling pathway diagrams Neurotransmission - receptor and channel signaling

GPCRs TYROSINE KINASES in urin GDNF Neurt Artem Persepin Glutamate Anandamide 1 2 3 4

2-AG α α α α uR

A/B* rin GFR GFR GFR GFR mGL CB1/2

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P in PI3K C PLCβ/γ PKC nNOS γ 3 FAK ER NFkB PDEI PKA RhoA RasGAP Src 2+ PKK2 Calcineurin Ca ROCK ERK FRS2 released Grb2 actin Src SOS RAS P CREB

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Neuroscience signaling pathways Cytoskeleton dynamics, neurite ASK1 P P extension CREB CREB MKK3/6 Other GPCRs ACh (Muscarinic): M1, M2, M3, M4 CRE p38MAPK Adrenalin: Alpha1b, 1c, 1d

ATP: P2Y MSK1 Dopamine: D1, D4 GABA: GABA receptor B P P

Opioid: μ, δ, κ CREB CREB Neurotensin: 1, 2 Neurokinin: NKA, NKB, NK1, Calcitonin * forward/reverse signalling Serotonin: 5HT-1B, D, E, F, 5HT-2A, B, 5HT-3, 4, 6, 7 inhibitory pathway CRE

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Src PSD95 Rap Shc Syntr K PL sin PI3 C Na+ Ca2+ Ca2+ γ Grb2 oph

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AKT/PKB Raf JNK nNOS SynGAP

MEK 1/2 P Ser133 Inhibition Cl- Cell survival CREB of signaling ERK 1/2

P P ELK RSK NFκB CRE CRE gene Long term B B transcription synaptic Bad CRE plasticity (LTP)

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Customer Service Tel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM) 87 3 ils p5 25 ls lear R gets 5-p EF2 ofibr SP16 fibri tar Nuc α Cdk Prot NMDA e.g M ation Cdk5 p25 nked uli cosyl O-li ic tim 2+ gly u pains th s th Ca gets plasm Cal tar e.g ta Dea l val Cyto 2 Cel survi Ub Ub Ub Ub Ub Ub Sp2 Ring 2 α p35 h7 Cdk5 Ub Ubc c-Abl P E2 IBR CAK ATP Ub h8 Ubc Ub Ub Ub Ring 1 Ub Ub E1 depletion) and the activation of programmed cell death machinery are also Ub )

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(Park Ub Ub Synphilin-1 Ub cdcrel-1 UBL Dorfin CHIP Ub Synaptotagmin substrates Ub RG Ub icity Cyclin E tubulins p38 PAC Tox Lewy body PGP9.5 aggregation UPS UCHL1 protein Neuroscience signaling pathways degraded dysfunction mitochondrial protein DA misfolded s Disease DA oxidation Dopamine DA -synuclein α P S 1 PINK-1 Genetic Mutation stress An abnormal increase in synuclein levels leads to the formation of protofibrils and insoluble fibrils, followed by proteasomal impairment, a process that cannot PINK- Oxidative mitochondrial DJ-1 DNA polymorphisms DNA echnical help - Neuroscience signaling pathway diagrams DJ-1 T Pathways of Parkinson’ Mutations in genes such as Parkin, DJ-1, synuclein, UCHL1, and PINK1 lead to disorders that variably resemble Parkinson's disease (PD). Dorfin SIAH are E3 ubiquitin ligases linked to PD (substrates unknown). DJ-1 and PINK1 appear protect against mitochondrial damage; functionally regulates mitochondria via phosphorylation (specific substrates unknown); DJ-1 by conversion to an acidic isoform after oxidative or environmental stress and translocation the outer mitochondrial membrane. be prevented in the absence of functional Parkin. Oxidative stress, energy crisis (i.e., believed to be factors that trigger the death of dopaminergic neurons in PD.

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5HT2C Receptor antibody ab23580 NRG1 type II antibody ab23314 ALDH1A1 antibody ab23375 NRG1 type III antibody ab23248 Arg3.1 antibody ab23382 Nudel antibody ab23550 BRN3A antibody ab23579 PAX2 antibody ab23788 Dbx1 antibody ab24605 PEN2 antibody ab18189 Delta-like 1 antibody ab23684 Piccolo antibody ab20664 Deltex antibody ab23665 PINK1 antibody ab23518 DLX 1 antibody ab23804 PSD95 antibody ab18258 DLX2 antibody ab18188 Robo3 antibody ab23562 DLX5 antibody ab23811 Semaphorin 3c antibody ab23575 DNER antibody ab23682 Semaphorin 7a antibody ab23578 Doublecortin (phospho S28) antibody ab23544 Slit3 antibody ab11018 Doublecortin (phospho S47) antibody ab23543 Synaptojanin antibody ab19904 Doublecortin (phospho S297) antibody ab23542 Syntenin antibody ab19903 EMX2 antibody ab23528 TBR1 antibody ab23361 Hippocalcin antibody ab24560 TBR2 antibody ab23345 KCNQ2 antibody ab22897 Tbx6 antibody ab23716 Lingo1 antibody ab23631 TPH2 antibody ab17733 Mesp2 antibody ab23733 TRPA1 antibody ab17734 Neurogenin 1 antibody ab23519 USP9x antibody ab19879 Nicotinic Acetylcholine Receptor alpha 7 antibody ab23832

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Target index Protein groupings Page Protein groupings Page Protein groupings Page 14-3-3 protein ...... 14,21,40 CD132 ...... 33 GAD ...... 48 5HT ...... 25,50,56 CD133 ...... 33 Galectin ...... 10 Acetylcholine ...... 47,53 CD24 ...... 33 GALR ...... 28,55 Acetylcholinesterase ...... 47 CD45 ...... 32 GAP ...... 36,38,45 Acrolein ...... 19 Cdk ...... 11 GCNF ...... 16 ACTH ...... 27,50 Cellubrevin ...... 40 GDNF ...... 12,13 arget index

T ADAM ...... 13,18,19 CGRP ...... 50 Gemin ...... 16 Adenosine Receptor ...... 55 Choline acetyltransferase ...... 38,47 GFAP ...... 6,31 Adiponectin ...... 28,29 CLACP ...... 18 Ghrelin ...... 28 Adrenergic Receptor ...... 54 Clathrin ...... 60 GHRHR ...... 26,55 Aggrecan ...... 11,33 CNPase ...... 32 GJB ...... 58 Agrin ...... 8,14,36 CNTF ...... 12,13,33,35 Gli ...... 16 AKT ...... 14,15 Coilin ...... 38 Glutamate ...... 47,48,54,61 ALDH1A1 ...... 22 COP ...... 45 Glutamate Receptor ...... 48,54 ALK ...... 37,45 Corticoliberin ...... 27,50 Glycine ...... 48,72 Als ...... 19,25 Corticotropin Releasing Factor ...... 27,50,55 Glycine Receptor ...... 48,57 AMIGO ...... 11 CPEB ...... 40 GRM ...... 54 Amisyn ...... 8,60 CRF ...... 27,32 GSK ...... 21 Amitriptyline ...... 53,62 Crk ...... 15,45 HAP40 ...... 22 Amphetamine ...... 53 CRMP ...... 11,25,36 Heparan sulfate proteoglycan ...... 9 AMPK ...... 18 CSN ...... 15,45,46 HIPPI ...... 22 Amyloid precursor protein ...... 18 CSPS ...... 49,61 HMG ...... 23,24 APH ...... 13,18,22 Ctip ...... 15,16 Homer ...... 40,54 Apolipoprotein ...... 19 CysLT ...... 55 HRH ...... 55 APXL ...... 59,63 Dab ...... 14,78 Humanin ...... 19 Artemin ...... 12 DARPP ...... 54 HUNK ...... 46 ASIC ...... 58,59,62 DDR ...... 58 Huntingtin ...... 22 Ataxin ...... 37 Dehydroepiandrosterone ...... 27 IL6 ...... 28,29 Attractin ...... 28 DHPR ...... 43 Insulin ...... 28 AVPR ...... 30,55 Dispatched ...... 16 Integrin ...... 34 Axotrophin ...... 11,62 DLL ...... 13 Internexin ...... 6 BACE ...... 20 Dopamine ...... 49,54,61 Jagged ...... 13,14 BAI ...... 36 DORFIN ...... 22,23 Ketanserin ...... 53,56 BDNF ...... 12,13 Doublecortin ...... 7,36,38 Kv ...... 59 Bestrophin ...... 8,63 Drebrin ...... 8,19,35,36 L1CAM ...... 11 beta Amyloid ...... 18 Dynamin ...... 40 Laminin ...... 6,8,24,25 BMP ...... 12,33 DYRK ...... 11 LANGF ...... 12 BNP ...... 50,51 EDG ...... 55 LAR ...... 29,40 C3 ...... 40,44 ELAV ...... 16,38 Leptin ...... 29 CACNB ...... 43 EMX ...... 16,33 LIM kinase ...... 16 Calbindin ...... 43 Endomorphin ...... 52 LIS ...... 16 Calcitonin ...... 55 ENSA ...... 26,58 LNX ...... 14 Calcium channel L type ...... 43 Ephrins ...... 11,36,45,46,59 LPHN ...... 55 Calpain ...... 25 ERAB ...... 19 MAML ...... 14 Calpastatin ...... 19,40,60 ERK ...... 21 MAP ...... 7,16,35,36,38 CaMK ...... 44 Estradiol ...... 26,50 MASS ...... 55 Cannabinoid Receptor ...... 54 Estrogen ...... 26,27 MC4 ...... 27,29,55 CAPON ...... 44,53 Exendin ...... 50 Melanopsin ...... 63 Carboxypeptidase ...... 28 Fatty Acid Synthase ...... 28 Melatonin ...... 27,29,55 CASK ...... 40,45 Fe65 ...... 19 Metabotropic Glutamate Receptor ...... 54 Caspase ...... 19 Fetuin ...... 8,16 Mint ...... 8,19,40,60 Cathepsin ...... 10,20 FMR ...... 16,51 Morphine ...... 52 CCKAR ...... 19,22,28 FOX ...... 12,16 MOSP ...... 10,32 CCKBR ...... 27,55 Fyn ...... 19,20,24 Munc ...... 60 CD11 ...... 32 GABA ...... 48,55,57,61 Muscarinic Acetylcholine Receptor ...... 53,56

Ordering information: www.abcam.com | Tech support: www.abcam.com/technical 92 Neuroscience resources: www.abcam.com/neuroscience Protein groupings Page Protein groupings Page Protein groupings Page Myelin ...... 10,32,33 Phosphotyrosine phosphatase ...... 12,29,39 Tau ...... 21,22

MYRIP ...... 7,40,60 PHOX ...... 16 Taurine ...... 48 T arget index NCAM ...... 8,16,36,37 Pin ...... 21 Tenascin ...... 9,17 NCS ...... 43 PINK1 ...... 23 Thrombospondin ...... 7 Nestin ...... 6,34,38 Pleiotrophin ...... 13 Thyroid Hormone ...... 30 Neurabin ...... 45 Plexin ...... 39 TRAF ...... 17 Neuregulin ...... 11,13,16,33 PMP ...... 10,33 TRAR ...... 58 Neurofigromin ...... 45 PP2 ...... 46 Trk ...... 13 Neurofilament ...... 6,7,37 PPP ...... 46 TRPM ...... 58,62 Neuroglobin ...... 16 Prealbumin ...... 21 Tryptamine ...... 48,49 Neurogranin ...... 44,46 Presenilin ...... 14,20 Tryptophan ...... 49 Neurokinin ...... 50,52,55 Prion protein PrP ...... 24 TSH ...... 30 Neuropeptide ...... 29,50-52,62 ProDynorphin ...... 52 Tubulin ...... 7,25,33,34,36,38 Neuropilin ...... 11 Progesterone ...... 27 Tyramine ...... 48,49 Neuroserpin ...... 11,25,37 Prohormone Convertase ...... 29 Ubiquitin ...... 20,22,25 Neurotrophin ...... 12,13 PROX ...... 16 UCP ...... 29 Neurturin ...... 13 PSD ...... 40,41,44,45,62 VAMP ...... 42,61 NGF ...... 13 Rab ...... 41,60 Vanilloid ...... 58,62,63 Nicastrin ...... 8,20 RAGE ...... 19,63 Vasopressin ...... 30,51 Nicotinic Acetylcholine Receptor ...... 57,91 RAI ...... 16 VCAM ...... 17 Nitrotyrosine ...... 52 Rapsyn ...... 58 VCP ...... 24 NMDAR ...... 58 Reelin ...... 8,9,11 VGLUT ...... 53,61 NMUR ...... 27,29,55,62 RIT ...... 63 Vimentin ...... 34,35,37 Nociception ...... 62,63 Robo ...... 11,37 VIP ...... 51 Noradrenaline ...... 49 ROR ...... 15 VRL ...... 51,58,63 NOS ...... 53,54 Ryanodine ...... 44 Zic ...... 17 Notch ...... 13,14 S100 ...... 31,32,43 NPFF ...... 51,52,55,62 SAP ...... 36,41,45 NR2E3 ...... 16 SCP ...... 16 NRP ...... 11,37 Serotonin ...... 50,56 NSE ...... 39 SHANK ...... 45 NUMB ...... 14 SHC ...... 19 Oct ...... 33 SIAH ...... 14,23 Olfactory Receptor ...... 63 SIRP ...... 8,17,41 Opioid ...... 52,56 Slit ...... 11,17 Orexin ...... 29,50,51 SNAP ...... 41,60,61 Oxytocin ...... 30,50,51 SOX ...... 17,34,39 P2X ...... 58 Spinesin ...... 10 P2Y ...... 55 SPON ...... 13 p35 ...... 37,46 Sprouty ...... 17 pan macrophage ...... 32 Stathmin ...... 12 Parathyroid Hormone ...... 30,55 STEP ...... 39 PARK7 ...... 23 Substance P ...... 51,52 Parkin ...... 23 Survivin ...... 33,39 Parvalbumin ...... 43 Synapsin ...... 41,60,61 PAX ...... 16,34 Synaptobrevin ...... 41,61 75 new PBP ...... 47,51 Synaptophysin ...... 41,42,61 Neuroscience PDE ...... 63 Synaptotagmin ...... 9,42,60 antibodies added Peripherin ...... 7 SynCAM ...... 9,17,42 Peropsin ...... 63 Syndecan ...... 42 to our catalog PGP9.5 ...... 23 SynGAP ...... 45 every month! PHF2 ...... 20 Synphilin ...... 23 Phosphoserine ...... 47 Syntaxin ...... 42,61 Phosphoserine/threonine ...... 46 Syntrophin ...... 9,58 Phosphotyrosine ...... 46,47 Synuclein ...... 20,23,24

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