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Biotium/Apoptotic, Necrotic, and Healthy Cells Quantification Kit/30018/50-assays

Thekitallowssimultaneousquantificationofapoptotic(green),necrotic(red)andallcells(blue)withinthesamecellpopulationbyflowcytometryorfluorescencemicroscopy.

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.

References

1.PhysicalBIOLOGy(2011)doi:10.1088/1478-3975/8/1/015006

2.PharmacologicalReports63,139(2011).

3.InternationalJournalofMolecularSciences9,56(2008)

.

4.Science300,91(2003).

5.JNeurosciMethods.36,229(1991).

6.FEBSLett203,99(1986).

7.Biosci.Rep.7,289(1987).

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