| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 490/520 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellAnalysis CellCytotoxicity |
| Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
- Treatcellswithtestcompoundsforadesiredperiodoftime(4-6hoursforJurkatcellstreatedwithcamptothecin)toinduceapoptosis.
Note:AnnexinVflowcytometricanalysisonadherentcellsisnotroutinelytestedsincespecificmembranedamagemayoccurduringcelldetachmentorharvesting.However,methodsforutilizingAnnexinVforflowcytometryonadherentcelltypeshavebeenpreviouslyreportedbyCasiola-Rosenetal.andvanEngelendetal(seeRefs1and2). - Centrifugethecellstoget2-5x105cells/tube.
- ResUSPendcellsin200µLofAssayBuffer(ComponentB).
- Add2µLofAnnexinV-FITC(ComponentA)intothecells.
Optional:Add2µLof100XPropidiumIodide(ComponentC)fornecrosiscells. - Incubateatroomtemperaturefor30to60minutes,protectedfromlight.
Optional:add200to300µLofAssayBuffer(ComponentB)toincreasevolumebeforeanalyzingthecellswithaflowcytometer(SeeStep7). - MonitorthefluorescenceintensityofAnnexinV-FITCusingtheFL1channel(Ex/Em=490/525nm),andmeasurethecellviABIlitywithpropidiumiodideusingtheFL2channel.
| 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.
