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Biotium/Apoptosis and Necrosis Quantification Kit/30017/50-assays

TheApoptosis&NecrosisQuantificationKitprovidesaconvenientassayforquantifyingapoptotic(green)andnecrotic(red)cellswithinthesamecellpopulationbyflowcytometryorfluorescencemicroscopy.

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).

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