Apoptosisandnecrosisaretwomajorprocessesbywhichcellsdie.Apoptosisisanactive,geneticallyregulateddisassemblyofthecellfromwithin.Duringapoptosis,phosphatidylserine(PS)istranslocatedfromtheinnertotheoutersurfaceofthecellforphagocyticcellrecognition.ThehumananticoagulantAnnexinVisa35kDa,Ca2-dependentphospholipid-bindingproteinwithahighaffinityforPS.AnnexinVlabeledwithfluorescein(FITC)(λEx/λEm=492/514nm)canidentifyapoptoticcellsingreenbybindingtoPSexposedontheouterleaflet.Necrosisnormallyresultsfromaseverecellularinsult,resultinginlossofplasmamembraneintegrity.EthidiumhomodimerIII(EthD-III)isahighlypositivelychargednucleicacidprobe,whichisimpermeanttolivecellsorapoptoticcells,butstainsnecroticcellswithredfluorescence(λEx/λEm(withDNA)=528/617nm).EthD-IIIisasuperioralternativetopropidiumiodide(PI)orethidiumhomodimerIbecauseofitssignificantlyhigheraffinityforDNAandhigherfluorescencequantumyield.TheApoptosis&NecrosisQuantificationKitprovidesaconvenientassayforquantifyingapoptotic(green)andnecrotic(red)cellswithinthesamecellpopulationbyflowcytometryorfluorescencemicroscopy.
KitComponents:
- FITC-AnnexinV
- EthidiumhomodimerIII
- 5Xbindingbuffer
Features:
- Simple:Simultaneousdetectionofbothapoptoticandnecroticcellsinasingleassay
- Fast:Asingle15-30minstainingstep
- Sensitive:EthD-IIIisasuperioralternativetobothPIandEthD-Iforstainingnecroticcells.
AlsoseetheApoptosisandNecrosisQuantitationKitPlus(catalogno.30065)andApoptotic,Necrotic,andHealthyCellsQuantitationKitPlus(catalogno.30066),whichfeatureAnnexinVconjugatedtoBiotium’ssuperiorgreenfluorescentdyeCF™488A,whichisbrighterandmorephotostablethanFITC.
References
1.BMCImmunology(2010)doi:10.1186/1471-2172-10-29.
2.BiosciRep7,289(1987).
3.FEBSLett203,99(1986).
