| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 565/575 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | R/L |
| Category | CellAnalysis CellCytotoxicity |
| Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1. Treatcellswithtestcompoundsforadesiredperiodoftime(4-6hoursforJurkatcellstreatedwithcamptothecin)toinduceapoptosis.
a. Note:AnnexinVflowcytometricanalysisonadherentcellsisnotroutinelytestedsincespecificmembranedamagemayoccurduringcelldetachmentorharvesting.However,methodsforutilizingAnnexinVforflowcytometryonadherentcelltypeshavebeenpreviouslyreportedbyCasiola-Rosenetal.andvanEngelendetal(seeRefs1and2).
2. Centrifugethecellstoget1-5×105cells/tube.
3. ResUSPendcellsin200μLofAssayBuffer(ComponentB).
4. ResuspendtheRPE-AnnexinV(ComponentA)with200μLPBSwith0.2%BSAtohave100Xstocksolution.
a. Note:Storethereconstitutedstocksolutionat4oC.DoNotFreeze
5. Add2μLof100XRPE-AnnexinVstocksolution(fromStep4)intothecells.
a. Optional:Add2µLof100XNuclearRed™DCS(ComponentC)fornecrosiscells.
6. Incubateatroomtemperaturefor20to60minutes,protectedfromlight.
7. Optional:add200to300μLofAssayBuffer(ComponentB)toincreasevolumebeforeanalyzingthecellswithaflowcytometer(SeeStep7).
8. MonitorthefluorescenceintensityofRPE-AnnexinVusingtheFL2channel(Ex/Em=488/585nm),andmeasurethecellviABIlitywithNuclearRed™DCSusingtheFL4channel.
| References&Citations |  PrinterFriendlyVersion |
1. BaeON,LimKM,NohJY,ChungSM,KimSH,ChungJH.(2009)Trivalentmethylatedarsenical-inducedphosphatidylserineexposureandapoptosisinplateletsmayleadtoincreasedthrombusformation.ToxicolApplPharmacol,239,144.
2. BurteaC,LaurentS,LancelotE,BalletS,MurariuO,RousseauxO,PortM,VanderElstL,CorotC,MullerRN.(2009)PeptidictargetingofphosphatidylserinefortheMRIdetectionofapoptosisinatheroscleroticplaques.MolPharm,6,1903.
3. DongHP,HolthA,KleinbergL,RuudMG,ElstrandMB,TropeCG,DavidsonB,RisbergB.(2009)Evaluationofcellsurfaceexpressionofphosphatidylserineinovariancarcinomaeffusionsusingtheannexin-V/7-AADassay:clinicalrelevanceandcomparisonwithotherapoptosisparameters.AmJClinPathol,132,756.
4. LiuT,ZhuW,YangX,ChenL,YangR,HuaZ,LiG.(2009)DetectionofapoptosisbasedontheinteractionbetweenannexinVandphosphatidylserine.AnalChem,81,2410.
5. MirnikjooB,BalasubramanianK,SchroitAJ.(2009)Suicidalmembranerepairregulatesphosphatidylserineexternalizationduringapoptosis.JBiolChem,284,22512.
6. MirnikjooB,BalasubramanianK,SchroitAJ.(2009)Mobilizationoflysosomalcalciumregulatestheexternalizationofphosphatidylserineduringapoptosis.JBiolChem,284,6918.
7. ThapaN,KimS,SoIS,LeeBH,KwonIC,ChoiK,KimIS.(2008)Discoveryofaphosphatidylserine-recognizingpeptideanditsutilityinmolecularimagingoftumourapoptosis.JCellMolMed,12,1649.
8. AiresV,HichamiA,FilomenkoR,PleA,RebeC,BettaiebA,KhanNA.(2007)Docosahexaenoicacidinducesincreasesin[Ca2+]iviainositol1,4,5-triphosphateproductionandactivatesproteinkinaseCgammaand-deltaviaphosphatidylserinebindingsite:implicationinapoptosisinU937cells.MolPharmacol,72,1545.
9. BalasubramanianK,MirnikjooB,SchroitAJ.(2007)Regulatedexternalizationofphosphatidylserineatthecellsurface:implicationsforapoptosis.JBiolChem,282,18357.
10. CauchonN,LangloisR,RousseauJA,TessierG,CadoretteJ,LecomteR,HuntingDJ,PavanRA,ZeislerSK,vanLierJE.(2007)PETimagingofapoptosiswith(64)Cu-labeledstreptavidinfollowingpretargetingofphosphatidylserinewithbiotinylatedannexin-V.EurJNuclMedMolImaging,34,247.
