| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 403/454 |
| MW | 555.94 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | EnzymeDetection PeptidasesandProteases |
| Related | CellApoptosis ApoptosisandCytotoxicity BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1. GeneralSolutionCaspaseAssaysUsingAMC,AFC,pNA,R110andProRedSubstrates
1.1. Preparea10mMstocksolutioninDMSO.
1.2. Preparea2Xcaspase substrate(50µM)assaysolutionasthefollowing:
50µLsubstratestocksolution
100 µLDTT(1M)
400 µLEDTA(100mM)
10mLTrisBuffer(20mM),pH=7.4
1.3. Mixequalvolumeofthecaspasestandardsorsampleswith2Xcaspasesubstrateassaysolution(fromStep1.2),andincubatethesolutionsatroomtemperatureforatleast1hour.
1.4. Monitorthefluorescenceusingafluorescencemicroplatereader,orabsorbanceusinganabsorbancemicroplatereader.
2. CellCaspaseAssaysUsingCell-PermeableFMKCaspaseProbes
2.1. Preparea2-5mMstocksolutioninDMSO.
2.2. Treatcellsasdesired.
2.3. Preparea2Xpermeablecaspasesubstrate(20µM)assaysolutionbydilutingtheDMSOstocksolution(fromStep2.1)inHankswith20mMHepesbuffer(HHBS).
2.4. Mixequalvolumeofthetreatedcellswith2Xcaspasesubstrateassaysolution(fromStep2.3),andincubatethecellsina37°C,5%CO2incubatorforatleast1hour.
2.5. WashthecellswithHHBSforatleastonce.
2.6. Monitorthefluorescenceintensitybyaflowcytometer,afluorescencemicroscopeorafluorescencemicroplatereader.
3. CellCaspaseAssaysUsingCell-PermeableFMKCaspaseProbes(For#13470-13476only)
3.1. Preparea250Xstocksolutionbyadding50µLDMSOintothevial.
3.2. Treatcellsasdesired.
3.3. Add250XDMSOstocksolution(fromStep3.1)intothecellsolutionata1:250ratio(suchas2uLto500uLcells),andincubatethecellsina37°C,5%CO2incubatorfor1hour.
3.4. WashthecellswithHHBSforatleastonce.
3.5. Monitorthefluorescenceintensitybyflowcytometer,fluorescencemicroscopyorfluorescentmicroplatereader.
| References&Citations |  PrinterFriendlyVersion |
1. LawrenceCP,ChowSC.(2012)SuppressionofhumanTcellproliferationbythecaspaseinhibitors,z-VAD-FMKandz-IETD-FMKisindependentoftheircaspaseinhibitionproperties.ToxicolApplPharmacol.
2. LiYY,YanCL.(2012)[InhibitionofelicitationofallergiccontactdermatitisbytopicaluseofZ-VAD-FMK,abroadcaspaseinhibitor:experimentinmice].ZhonghuaYiXueZaZhi,92,1992.
3. MeslinB,BeavoguiAH,FaselN,PicotS.(2011)PlasmodiumfalciparummetacaspasePfMCA-1triggersaz-VAD-fmkinhibitableproteasetopromotecelldeath.PLoSOne,6,e23867.
4. MullerI,LamersMB,RitchieAJ,DominguezC,Munoz-SanjuanI,KiselyovA.(2011)Structureofhumancaspase-6incomplexwithZ-VAD-FMK:Newpeptidebindingmodeobservedforthenon-canonicalcaspaseconformation.BioorgMedChemLett,21,5244.
5. AbaamraneL,RaffinF,SchmerberS,SendowskiI.(2011)Intracochlearperfusionofleupeptinandz-VAD-FMK:influenceofantiapoptoticagentsongunshot-inducedhearingloss.EurArchOtorhinolaryngol,268,987.
6. EquilsO,Moffatt-BlueC,IshikawaTO,SimmonsCF,IlievskiV,HirschE.(2009)Pretreatmentwithpancaspaseinhibitor(Z-VAD-FMK)delaysbutdoesnotpreventintraperitonealheat-killedgroupBStreptococcus-inducedpretermdeliveryinapregnantmousemodel.InfectDisObstetGynecol,2009,749432.
7. LiYY,YanCL,XuW.(2008)[AttenuationofallergiccontactdermatitisbyZ-VAD-FMK,abroadcaspaseinhibitor:experimentwithmice].ZhonghuaYiXueZaZhi,88,3153.
8. WitteMD,DescalsCV,deLavoirSV,FloreaBI,vanderMarelGA,OverkleeftHS.(2007)Bodipy-VAD-Fmk,ausefultooltostudyyeastpeptideN-glycanaseactivity.OrgBiomolChem,5,3690.
9. HengBC,ClementMV,CaoT.(2007)CaspaseinhibitorZ-VAD-FMKenhancesthefreeze-thawsurvivalrateofhumanembryonicstemcells.BiosciRep,27,257.
10. VejuxA,LizardG,TourneurY,RiedingerJM,FrouinF,KahnE.(2007)Effectsofcaspaseinhibitors(z-VAD-fmk,z-VDVAD-fmk)onNileRedfluorescencepatternin7-ketocholesterol-treatedcells:investigationbyflowcytometryandspectralimagingmicroscopy.CytometryA,71,550.
