Apoptosisandnecrosisaretwomajorprocessesbywhichcellsdie.Apoptosisisanactive,geneticallyregulateddisassemblyofthecellfromwithin.Duringapoptosis,phosphatidylserine(PS)istranslocatedfromtheinnertotheouterleafletoftheplasmamembraneforphagocyticcellrecognition.ThehumananticoagulantAnnexinVisa35kDa,Ca2-dependentphospholipid-bindingproteinwithhighaffinityforPS.AnnexinVlabeledwithCF™488A(Abs/Em:490/515nm)stainsapoptoticcellsgreenbybindingtoPSexposedonthecellsurface.CF™488Aisspectrallysimilartofluorescein(FITC),withmuchbrighterandmorephotostablefluorescence.
Necrosisnormallyresultsfromseverecellularinsult.Bothinternalorganelleandplasmamembraneintegrityarelost,resultinginthereleaseofcellularcontentsintothesurroundingenvironment.EthidiumhomodimerIII(EthD-III)isahighlypositivelychargednucleicacidprobe,whichisimpermeanttolivecells,butstainscellswithcompromisedmembraneintegritywithredfluorescence(Abs/Em(withDNA):528/617nm).EthD-IIIisasuperioralternativetopropidiumiodide(PI)orethidiumhomodimerIbecauseofitssignificantlyhigheraffinityforDNAandhigherfluorescencequantumyield.TheApoptosis&NecrosisQuantitationKitPlusprovidesaconvenientassayforquantifyingapoptotic(green)andnecrotic(red)cellswithinthesamecellpopulationbyflowcytometryorfluorescencemicroscopy.
KitComponents:
- CF™488A-AnnexinV
- EthidiumhomodimerIII
- 5Xbindingbuffer
Features:
- Simple:Simultaneousdetectionofbothapoptoticandnecroticcellsinasingleassay
- Fast:Asingle15-30minstainingstep
- Sensitive:CF™488A-AnnexinVisbrighterandmorephotostablethanFITC-AnnexinV,whileEthD-IIIisasuperioralternativetobothPIandEthD-Iforstainingnecroticcells.
References
1.Cvjeticanin,T.,etal.TCellsCooperatewithPalmiticAcidinInductionofBetaCellApoptosis.BMCImmunologydoi:10.1186/1471-2172-10-29(2010).
2. Biol.ofReproduction(2016),doi:10.1095/biolreprod.115.134924
