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Advanced Targeting Systems Product Catalog AAddvvaanncceedd TTaarrggeettiinngg SSyysstteemmss PPrroodduucctt CCaattaalloogg Molecular Surgery for Scientists (877) 889-2288 toll-free 10451 Roselle Street, #300 (858) 642-1988 telephone www.ATSbio.com website San Diego, CA 92121 USA (858) 642-1988 fax [email protected] email Table of Contents Saporin ...........................................................5 Secondary Conjugates ................................21 Saporin ............................................................................5 Anti-M-ZAP (primary mouse monoclonal IgM )..............21 Anti-Saporin ....................................................................5 Fab-ZAP (primary mouse monoclonal IgG , monovalent )22 Targeted Toxins .............................................6 Goat-ZAP (primary goat monoclonal IgG ) ....................22 Hu g-M -ZAP (primary human monoclonal IgM ) .............23 192-IgG-SAP (rat NGFr ) .................................................6 Hum-ZAP (primary : human monoclonal IgG ) ...............23 Acetylated LDL-SAP (rat microglia ) ...............................7 Mab-ZAP (primary mouse monoclonal IgG ) .................24 Anti-CD22-SA P (human CD22 ) ......................................7 Anti-CD25-SAP (human CD25 ) .....................................8 Rab-ZAP (primary rabbit polyclonal IgG )......................24 Anti-CD25-SAP (mouse CD25 ) .....................................8 Rat-ZAP (primary rat monoclonal IgG ) .........................25 Anti-CD103-SAP (CD103) ...............................................9 Custom Conjugates ......................................................25 Anti-DAT-SAP (rat / human dopamine transporter ........9 Streptavidin-ZAP (biotinylated material ) .......................26 Anti-DBH-SAP (r at dopamine beta-hydroxylase )..........10 Biotinylation Service ......................................................26 Anti-SERT-SAP (rat / human SERT) .............................11 Bombesin-SAP (GRP receptor ).....................................11 Control Conjugates .....................................27 CTB-SAP (GM 1 Ganglioside ) .......................................12 Blank-SAP (for peptide conjugates )..............................27 CRF-SAP (CRF receptor 1 ) ..........................................12 Goat IgG-SAP ...............................................................28 CCK-SAP (CCKr1 and CCKr2 ).....................................12 Human IgG-SAP ...........................................................28 Dermorphin-SAP / MOR-SAP (MOR ) ...........................13 Mouse IgG-SAP ............................................................29 FGF-SAP (FGF receptor s) ............................................13 Mouse IgM-SAP ............................................................29 GAT1-SAP (GABA-1 transporter, GAT-1 )......................14 Rabbit IgG-SAP ............................................................30 Galanin-SAP (galanin-1 receptor ).................................14 Rat IgG-SAP .................................................................30 IB4-SAP (a-D-Galacto pyranoside residues ) .................15 Fluorescent Conjugates ..............................31 Leptin-SAP (leptin receptor )..........................................15 Custom Fluorescent Labeling .......................................32 Mac-1-SAP (mouse / human CD11b receptor ) .............16 Antibodies ....................................................34 Mac-1-SAP (rat CD11b receptor )..................................16 ME20.4-SAP (mammalian NGFr ) .................................17 Proteins, Peptides & Antigens ...................46 Melanopsin-SAP (melanopsin , ipRGCs ).......................17 Index .............................................................59 mu p75-SAP (mouse NGFr )..........................................18 NPY-SAP (neuropeptide Y receptor ).............................18 Oxytocin-SAP (oxytocin receptor ).................................19 OX7-SAP (multi-species Thy-1.1 ).................................19 SSP-SAP (substance P receptor ) .................................20 SP-CTA (substance P receptor )....................................20 Table of Contents 2 The Technology 3 Saporin 4 Targeted Toxins 5 Secondary Conjugates 20 Control Conjugates 26 Fluorescent Conjugates 30 Antibodies 33 Proteins, Peptides & Antigens 45 Index 58 Page 3 The Technology Follow these simple illustrations to understand how this advanced targeting system can work for you. Or watch a multi-media presentation demonstrating this technology at www.ATSbio.com/movie 1) 2) 3) 4) Get an antibody specific to your cell type. Use a potent toxin that is ATS puts them The conjugate seeks together out target markers Or use anything safe in the lab, to make a powerful on the cell surface. that binds to a cell but lethal inside a cell. targeting agent. surface molecule and is internalized. 5) 6) 7) 8) Once attached to its target receptor, the The bond between The ribosomes are conjugate is internalized. The SAPORIN SAPORIN and the inactivated Cells which do not have conjugate enters the targeting agent is resulting in the target receptor cell. broken in the cell death. will not be affected. endosome. Pretty simple, huh? Now just imagine the targets you can eliminate in your research! Page 4 Saporin (pronounced ) Saporin (Cat. #PR-01: 100 µg) Saporin is a ribosome-inactivating protein, molecular weight 30 kDa, from seeds of the plant Saponaria officinalis . Effective Tool • An experimental control with targeted toxins • Safe for laboratory use under normal safety conditions • LD 50 in mice at 4 mg/kg • Does not have a method of cell entry on its own Sample protocol For most targeted toxins, there are control conjugates available (pp. 23-25); these are the most accurate controls to use. For a few products, saporin is used as control. Calculate amount of saporin needed on a molar basis. For example, FGF-SAP (Cat. #IT-38) has an average mol. wt. of 63 kDa, and 71% of that is saporin (approximately 1.5 molecules of saporin per molecule of cytokine). For injections of 5 µg of immunotoxin, an appropriate control quantity would be 3.6 µg. Anti-Saporin (Cat. #AB-15; AB-15 AP ; AB-15 hRP ; AB-17 AP , AB-41 AP ) Specificity AB-1 5Ap Goat Anti-Saporin, • goat anti-saporin, 100 µl (Cat. #AB-15) affinity purified • goat anti-saporin affinity purified, 50 µg (Cat. #AB-15 AP ) Lane 1: Molecular weight standards (Invitrogen SeeBlue) • goat anti-saporin HRP-labeled, 200 µg (Cat. #AB-15 HRP ) Lane 2: 50 ng of Saporin • chicken anti-saporin affinity purified, 50 µg (Cat. #AB-17 AP ) probed with Goat anti-Saporin • rabbit anti-saporin affinity-purified, 50 µg (Cat. #AB-41 AP ) affinity purified (AB-15 Ap ) at 1.0 µg/ml Applications Lane 3: 25 ng of Saporin • immunoblotting, ELISA, immunohistochemistry probed with Goat anti-Saporin affinity purified (AB-15 Ap ) at 1.0 AB-1 5hrp Goat Anti- µg/ml Saporin, hrp-labeled Saporin (200, 100, 50, and 25 ng) was run on a 10% SDS-pAGE gel AB-17 Ap Chicken and transferred to pVDF membrane. Anti-Saporin, The blot was blocked with 4% affinity-purified NFM/TBS, then incubated overnight Lane 1: Molecular with 0.5 µg/ml (Lanes 11-15), 1 µg/ml weight standards (Lanes 6-10), or 5 µg/ml (Lanes 1-5) (Invitrogen of antibody. The blot was washed SeeBlue) and developed with 4-chloro-1- naphthol and hydrogen peroxide. Lane 2: 100 ng of Saporin probed Lane 1, 6, 11: Molecular weight with AB-17 Ap at a standards (Invitrogen SeeBlue) concentration of Lane 2, 7, 12: 200 ng Saporin 0.5 µg/ml Lane 3, 8, 13: 100 ng Saporin Lane 4, 9, 14: 50 ng Saporin Page 5 Lane 5, 10, 15: 25 ng Saporin Targeted Toxins 192-IgG-SAP TARGET: rat NGFr a chemical conjugate of a mouse monoclonal antibody to rat p75 NTR and the ribosome-inactivating protein, saporin Intraventricular injection of 192-IgG-SAP results in almost complete elimination of low affinity nerve growth factor receptor (p75 NTR )-positive cells in the rat basal forebrain. This immunotoxin provides researchers with a powerful lesioning tool — more specific and effective than chemical, surgical or electrolytic lesioning. 192-IgG-SAP (conjugate alone) Cat # Units IT-01-25 25 micrograms IT-01-100 100 micrograms IT-01-250 250 micrograms Kit: conjugate and controls (saporin, control conjugate) CONTrOL TrEATED Cat # Units p75 NTR -positive neurons of rat cholinergic icv injection of 192-Saporin KIT-01-25 25 micrograms forebrain >95% elimination of target neurons KIT-01-100 100 micrograms Photos supplied by C. Wrenn, R.G. Wiley KIT-01-250 250 micrograms related products: Cy3-labeled 192-IgG (Cat. #FL-01), Alexa488-labeled 192-IgG (Cat. #FL-03), Mouse IgG-SAp (Cat. #IT-18), mu p75-SAp (Cat. #IT-16), ME20.4-SAp (Cat. #IT-15) Page 6 Acetylated LDL-SAP TARGET: rat microglia a chemical conjugate of acetylated human low density lipoprotein and the ribosome-inactivating protein, saporin Acetylated LDL-Saporin (acetylated LDL-SAP) has been used effectively to eliminate microglia from primary cultures of rat hippocampal neurons, while having no deleterious effect on neurons or astroglia. After 24 hours exposure, 3 µg/ml of acetylated-LDL-SAP selectively kills more than 90% of microglia cells. Acetylated LDL-SAP (conjugate alone) Cat # Units IT-08-25 25 micrograms IT-08-100 100 micrograms IT-08-250 250 micrograms Kit: conjugate and control (saporin) Cat # Units NR8383 cells are plated at 10 4 cells/well and incubated overnight. KIT-08-25 25 micrograms Saporin and acLDL-SAp are added in 10-µl volumes and the plates are KIT-08-100 100 micrograms
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