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.Thekitalsoincludesthemembrane-permeantbluefluorescentDNAdyeHoechst33342(λEx/λEm(withDNA)=350/461nm)forvisualizingallcells.Thekitallowssimultaneousquantificationofapoptotic(green),necrotic(red)andallcells(blue)withinthesamecellpopulationbyflowcytometryorfluorescencemicroscopy.
KitComponents:
- FITC-AnnexinV
- EthidiumHomodimerIII
- Hoechst33342
- 5XbindingBuffer
Features:
- Simple:Simultaneousdetectionofapoptotic,necroticandhealthycellsinasingleassay
- Fast:Asingle15-30minstainingstep
- Sensitive:EthD-IIIisasuperioralternativetobothPIandEthD-Iforstainingnecroticcells.
AlsoseetheApoptosisandNecrosisQuantitationKitPlus(catalogno.30065)andApoptotic,Necrotic,andHealthyCellsQuantitationKitPlus(catalogno.30066),whichfeatureAnnexinVconjugatedtoBiotium’ssuperiorgreenfluorescentdyeCF™488A,whichisbrighterandmorephotostablethanFITC.
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