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Roche/11684795910 / In Situ Cell Death Detection Kit, Fluorescein现货
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详细信息：InSituCellDeathDetectionKit,Fluorescein 现货供应Kitforthedetectionandquantificationofapoptosis(programmedcelldeath)atthesingle-celllevel,basedonlabelingofDNAstrandbreaks(TUNELtechnology):AnalysisbyfluorescencemicroscopyorflowcytometryApplicationQualitativedetectionofapoptosisatthesingle-celllevelbyfluorescencemicroscopyandquantitativedetectionbyflowcytometry.BenefitsProductDescriptionSamplematerial: CellsinsUSPension,cytospinandcellsmearpreparations,adherentcellsgrownonslides,andfrozenandparaffin-embeddedtissuesections.BackgroundInformationWidelyusedmethodstodetermineapoptosisincludetheanalysisofthegenomicDNAbyagarose-gelelectrophoresisandDNAfragmentationassaysbasedon 3H-thymidineand,alternatively,5-Bromo-2-deoxy-uridine.Themethodsinvolvetheseparationoffragmented,lowmolecularweightDNAfromunfragmented,highmolecularweightDNAinagivencellpopulation.Thus,thesemethodsdonotprovideinformationaboutthefateofanindividualcellinagivencellpopulation,orparticularly,intissuesections.Alternatively,individualapoptoticcellsmaybemicroscopicallyrecognizedbecauseofthecharacteristicappearanceofnuclearchromatincondensationandfragmentation,butthismethodissubjectiveandlimitedtoarelativelynarrowtimewindowwhenthemorphologicalchangesareatamaximum.ThehallmarkofapoptosisisDNAdegradation,whichinearlystages,isselectivetotheinternucleosomalDNAlinkerregions.TheDNAcleavagemayyielddouble-strandedandsingle-strandedDNAbreaks(nicks).Bothtypesofbreakscanbedetectedbylabelingthefree3-OHterminiwithmodifiednucleotides(e.g., biotin-dUTP,DIG-dUTP,fluorescein-dUTP)inanenzymaticreaction.Theenzymeterminaldeoxynucleotidyltransferase(TdT)catalyzesthetemplate-independentpolymerizationofdeoxyribonucleotidestothe3-endofsingle-anddouble-strandedDNA.ThismethodhasalsobeentermedTUNEL(TdT-mediateddUTP-X nick end labeling).Alternatively,free3-OHgroupsmaybelabeledusingDNApolymerasesbythetemplate-dependentmechanismcallednicktranslation.However,theTUNELmethodisconsideredtobemoresensitiveandfaster.ContentsPrincipleThe InSitu CellDeathDetectionKitFluorescein isbasedonthedetectionofsingle-anddouble-strandedDNAbreaksthatoccurattheearlystagesofapoptosis.ApoptoticcellsarefixedandpermeABIlized.Subsequently,thecellsareincubatedwiththeTUNELreactionmixturethatcontainsTdTandfluorescein-dUTP.Duringthisincubationperiod,TdTcatalyzestheadditionoffluorescein-dUTPatfree3-OHgroupsinsingle-anddouble-strandedDNA.Afterwashing,thelabelincorporatedatthedamagedsitesoftheDNAisvisualizedbyflowcytometryand/orfluorescencemicroscopy.Sensitive: Thedirectlabelingprocedureusingfluorescein-dUTPreducesbackgroundlabelingFast: Theuseoffluorescein-dUTPallowsanalysisofthesamplesdirectlyaftertheTUNELreactionConvenient: NosecondarydetectionsystemrequiredAccurate: Identificationofapoptosisatamolecularlevel(DNA-strandbreaks)andidentificationofcellsattheveryearlystagesofapoptosisEnzymeSolution(TdT),5vialsLabelSolution(fluorescein-dUTP),5vials
