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T E C H N Ic a L R E F E R E N Abbreviations A E α alpha ED50 effective dose (for 50% of effect) Ab antibody EDTA ethylenediaminetetraacetic acid Ac-DEVD-AMC acetyl-Asp-Glu-Val-Asp-7-amino-4-methyl coumarin EGF epidermal growth factor (fluorogenic substrate for caspase-3/7) EGFR epidermal growth factor receptor Ac-DEVD-CHO acetyl-Asp-Glu-Val-Asp-1-aldehyde EGTA ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (reversible aldehyde inhibitor of caspase-3/7) ELISA enzyme-linked immunosorbent assay Ac-DEVD-pNA acetyl-Asp-Glu-Val-Asp-pNA (colorimetric substrate for caspase-3/7) em emission Ac-YVAD-AMC acetyl-Tyr-Val-Ala-Asp-amino methyl coumarin ERK1, 2 extracellular signal-regulated protein kinase 1, 2 (fluorogenic substrate for caspase-1) EtBr ethidium bromide Ac-YVAD-CHO acetyl-Tyr-Val-Ala-Asp-1-aldehyde EtOH ethanol (reversible aldehyde inhibitor of caspase-1) ex excitation ADP adenosine diphosphate Ad-2 adenovirus-2 F AKAP A-kinase anchoring protein FAB/MS fast atomic bombardment mass spectrometry a.m.u. atomic mass unit FACS fluorescence-activated cell sorting AMC 7-amino-4-methyl coumarin FGF fibroblast growth factor amol attomole (10–18 mole) FITC fluorescein isothiocyanate AMP adenosine monophosphate FITC-VAD-FMK FITC-carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- AMV avian myeloblastosis virus fluoromethylketone (fluorescent marker for caspase activity) AP alkaline phosphatase FL fluorescein –15 AP1, 2 activator protein 1, 2 fmol femtomole (10 mole) APC film automatic processor-compatible film G ARE AU-rich element γ gamma ATP adenosine triphosphate g gram B GAPDH glyceraldehyde-3 phosphate dehydrogenase GDNF glial cell line-derived neurotrophic factor β beta BCIP 5-bromo-4-chloro-3-indolyl phosphate GFAP glial fibrillary acidic protein BDNF brain-derived neurotrophic factor GFP green fluorescent protein BMV brome mosaic virus GLB Glo Lysis Buffer bp base pairs GMO genetically modified organism BRET bioluminescence resonance energy transfer GMP guanosine monophosphate BSA bovine serum albumin GPDH glycerol 3-phosphate dehydrogenase BYDV barley yellow dwarf virus GQ genome qualified GST glutathione-S-transferase TECHNICAL REFERENCE C GTP guanosine triphosphate CaM KII calcium/calmodulin-dependent protein kinase II cAMP adenosine-3 ,5 -cyclic monophosphate (cyclic AMP) H ′ ′ 3 CAT chloramphenicol acetyltransferase H tritium CBZ benzyloxycarbonyl h human CCD charge-coupled device (camera) HC high concentration CCLR Cell Culture Lysis Reagent hCL1 synthetic CL1, a protein degradation signal cdc2 cell division cycle 2 protein HCV hepatitis C virus cDNA complementary DNA HDPE high-density polyethylene cfu colony forming unit HIV human immunodeficiency virus hluc+ codon-optimized firefly luciferase gene cGMP guanosine-3′,5′-cyclic monophosphate (cyclic GMP) Ci Curie hlucCP+ hluc+ with 3′ hCL1 and hPEST sequences CIAP calf intestinal alkaline phosphatase hlucP+ hluc+ with 3′ hPEST sequence CKI, CK-1 casein kinase I hPEST synthetic PEST, a protein degradation signal CKII, CK-2 casein kinase II HPLC high-pressure or high-performance liquid chromatography cm centimeter hRluc synthetic Renilla luciferase gene CMM canine pancreatic microsomal membranes hRlucCP hRluc with 3′ hCL1 and hPEST sequences CMV cytomegalovirus hRlucP hRluc with 3′ hPEST sequence CN 4-chloro-1-naphthol (a horseradish peroxidase substrate) HRP horseradish peroxidase CNBr cyanogen bromide HSV herpes simplex virus CNS central nervous system HTP high throughput CNTF ciliary neurotrophic factor HTS high-throughput screening CODIS COmbined DNA Index System I cpm counts per minute IC50 inhibitory concentration (50% inhibition) CPP32 caspase-3 (DEVDase) ICC immunocytochemistry CREB cAMP response element binding protein ICE interleukin-1β-converting enzyme (caspase-1) IGF insulin-like growth factor Ct cycle threshold CTP cytidine triphosphate IgG immunoglobulin G CXR carboxy-X-tetramethylrhodamine IgY immunoglobulin Y (chicken egg yolk immunoglobulin) D IHC immunohistochemistry Da daltons IL-4 interleukin-4 DAB diaminobenzidine IPTG isopropyl β-d-thiogalactopyranoside DAG diacylglycerol iso-dC isodeoxycytosine iso-dG isodeoxyguanosine DAPI 4′,6-diamidino-2-phenylindole DEPC diethyl pyrocarbonate IVEC in vitro expression cloning DEVDase caspase protease activity on the DEVD peptide IVT in vitro transcription ddRNAi DNA-directed RNA interference J diAcFAM diacetyl carboxyfluorescein JNK c-Jun N-terminal kinase DLR Dual-Luciferase® Reporter JOE 6-carboxy-4′,5′-dichloro-2′,7′-dimethoxy-fluorescein DMSO dimethyl sulfoxide K DNA deoxyribonucleic acid kb kilobase; kilobase pairs DNA-PK DNA-dependent protein kinase kDa kilodalton DNase deoxyribonuclease Km Michaelis-Menten constant dNTP deoxynucleotide triphosphate DOGS dioctadecylamidoglycyl spermine DOPE L-dioleoyl phosphatidylethanolamine DPPIV dipeptidyl peptidase IV dsDNA double-stranded DNA dsRNA double-stranded RNA DTT dithiothreitol Abbreviations L PPase protein phosphatase λ lambda PTPase protein tyrosine phosphatase L liter PTK protein tyrosine kinase LAR Luciferase Assay Reagent PTT protein truncation test LDH lactate dehydrogenase PVDF polyvinylidene fluoride LDPE low-density polyethylene PVP polyvinylpyrrolidone LNGFR low-affinity NGF receptor (p75 neurotrophin receptor) Q LSC liquid scintillation counting qPCR quantitative PCR luc native firefly luciferase gene qRT-PCR quantitative reverse transcriptase PCR luc+ synthetic firefly luciferase gene R LY 294002 2-(4-morpholinyl)-8-phenyl-4 H-1-benzopyran-4-one r recombinant (PI-3 kinase inhibitor) RAPD random amplified polymorphic DNA M RF DNA replicative form DNA m murine; milli RISC RNA-induced silencing complex M molar RLB Reporter Lysis Buffer mAb monoclonal antibody RLU relative light units MALDI-TOF matrix-assisted laser desorption/ionization-time of flight mass Rluc native Renilla luciferase gene spectrometry RNA ribonucleic acid MAO monoamine oxidase RNAi RNA interference MAPK mitogen-activated protein kinase RNase ribonuclease MCS multiple cloning site rNTP ribosylnucleotide triphosphate, (same as NTP) MEK MAPK kinase ROS reactive oxygen species –3 mg milligram (10 g) rRNA ribosomal RNA ® MGFP Monster Green Fluorescent Protein RT reverse transcriptase –6 µg microgram (10 g) RT-PCR reverse transcription PCR min minute µl microliter (10–6L) S µM micromolar (10–6M) SAM S-adenosylmethionine miRNA micro RNA SAM streptavidin matrix ml milliliter (10–3L) SAP shrimp alkaline phosphatase MLCK kinase 338, myosin light chain kinase SB 203580 (4-[4′-fluorophenyl]-2-[4′-methylsulfinylphenyl]-5-[4′-pyridyl] imidazole), a p38 MAP kinase inhibitor M-MLV Moloney murine leukemia virus SDS sodium dodecyl sulfate mm millimeter (10–3m) shRNA short hairpin RNA mM millimolar (10–3M) siRNA short interfering RNA TECHNICAL REFERENCEmRNA messenger RNA SNP single nucleotide polymorphism MTS 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2- (4-sulfophenyl)-2H-tetrazolium, inner salt SP1 specificity protein 1 MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide SSB single-stranded DNA binding protein MW molecular weight ssDNA single-stranded DNA St stearated N STR short tandem repeat N Normal SV40 simian virus 40 NaOAc sodium acetate NBT nitro blue tetrazolium T NED N-1-napthylethylenediamine dihydrochloride TAE tris acetate EDTA NF N-terminal fusion TAP tobacco acid pyrophosphatase TBE tris borate EDTA NF-κB nuclear factor kappa B NGF nerve growth factor TCA trichloroacetic acid nmol nanomole (10–9 mole) TdT terminal deoxynucleotidyl transferase NMR nuclear magnetic resonance TE Tris-EDTA buffer nt nucleotide TEMED N,N,N′,N′-tetramethylethylenediamine NT-3, NT-4 neurotrophin-3, neurotrophin-4 TFIIB transcription factor IIB TGF transforming growth factor 1, 2 NTP nucleotide triphosphate (same as rNTP) β1,2 β TK thymidine kinase O TLC thin-layer chromatography OCT1 octamer-binding transcription factor 1 Tm melting temperature ONPG o-nitrophenyl -d-galactopyranoside β TMB 3,3′,5,5′-tetramethylbenzidine P TMR carboxy-tetramethylrhodamine P450 cytochrome P450 TNFα tumor necrosis factor-α pAb polyclonal antibody TPA 12-O-tetradecanoylphorbol 13-acetate PAGE polyacrylamide gel electrophoresis TPCK N-p-tosyl-L-phenylalanine chloromethyl ketone PARP poly (ADP-ribose) polymerase Trk tyrosine kinase neurotrophin receptor family PCR polymerase chain reaction tRNA transfer RNA PD 98059 2′-amino-3′-methoxyflavone, MEK1 inhibitor TTP thymidine triphosphate PES phenazine ethosulfate TUNEL TdT-mediated dUTP nick-end labeling PEG polyethylene glycol U –12 pg picogram (10 g) u unit Pgp P-glycoprotein UTP uracil triphosphate Pi inorganic phosphate PKA cAMP-dependent protein kinase V PKC protein kinase C (v/v) volume:volume ratio PKG cGMP-dependent protein kinase VAChT vesicular acetylcholine transporter V maximum velocity (enzyme kinetics) PI 3-K phosphatidylinositol 3-kinase max PLB Passive Lysis Buffer W PMA phorbol 12-myristate 13-acetate (TPA), PKC activator (w/v) weight:volume ratio pmol picomole (10–12 mole) X PMP paramagnetic particle d X-Gal 5-bromo-4-chloro-3-indolyl-β- -galactopyranoside PMS phenazine methosulfate Z PMSF phenylmethylsulfonyl fluoride Z-DEVD-R110 bis-(N-CBZ-L-aspartyl-L-glutamyl-L-valyl-aspartic acid amide) pNA p-nitroaniline rhodamine 110 (fluorogenic substrate for caspase 3/7) PNGase F peptide N-glycosidase F Z-VAD-FMK carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone PNPP p-nitrophenyl phosphate (pan caspase inhibitor) Poly(A) polyadenylation sequence zmol zeptomole (10–21 mole) PART #GE624 PROMEGA CORPORATION 2800 WOODS HOLLOW ROAD MADISON, WI 53711-5399 USATELEPHONE 608-274-4330 www.promega.com ©2010 ALL RIGHTS RESERVED.
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