Phosphatidylserine(PS)istranslocatedfromtheinnertotheoutersurfaceofthecellforphagocyticcellrecognition.Thehumananticoagulant,AnnexinV,isa35kDaCa2-dependentphospholipidproteinwithahighaffinityforPS.AnnexinVlabeledwithCF™488A(Abs/Em=490/515nm)canidentifyapoptoticcellsingreenbybindingtoPSexposedontheouterleaflet.OurCF™488Adyeissuperiortofluorescein/FITCasitisnotaffectedbypHandhasmuchbetterphotostABIlity.Propidiumiodide(PI)isimpermeanttolivecellsorearlyapoptoticcells,butstainsnecroticcellsandlateapoptoticcellswithredfluorescence(Abs/EmwithDNA=535/617nm).BothPIandAnnexinV-CF™488Acanbeexcitedusingthe488nmlineofaflowcytometer.
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