详细信息：
InSituCellDeathDetectionKit,Fluorescein 现货供应
Kitforthedetectionandquantificationofapoptosis(programmedcelldeath)atthesingle-celllevel,basedonlabelingofDNAstrandbreaks(TUNELtechnology):Analysisbyfluorescencemicroscopyorflowcytometry
Application
Qualitativedetectionofapoptosisatthesingle-celllevelbyfluorescencemicroscopyandquantitativedetectionbyflowcytometry.
Benefits
ProductDescription
Samplematerial: CellsinsUSPension,cytospinandcellsmearpreparations,adherentcellsgrownonslides,andfrozenandparaffin-embeddedtissuesections.
BackgroundInformation
WidelyusedmethodstodetermineapoptosisincludetheanalysisofthegenomicDNAbyagarose-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.Contents
Principle
The InSitu CellDeathDetectionKitFluorescein isbasedonthedetectionofsingle-anddouble-strandedDNAbreaksthatoccurattheearlystagesofapoptosis.
ApoptoticcellsarefixedandpermeABIlized.Subsequently,thecellsareincubatedwiththeTUNELreactionmixturethatcontainsTdTandfluorescein-dUTP.Duringthisincubationperiod,TdTcatalyzestheadditionoffluorescein-dUTPatfree3-OHgroupsinsingle-anddouble-strandedDNA.Afterwashing,thelabelincorporatedatthedamagedsitesoftheDNAisvisualizedbyflowcytometryand/orfluorescencemicroscopy.Sensitive: Thedirectlabelingprocedureusingfluorescein-dUTPreducesbackgroundlabeling
Fast: Theuseoffluorescein-dUTPallowsanalysisofthesamplesdirectlyaftertheTUNELreaction
Convenient: Nosecondarydetectionsystemrequired
Accurate: Identificationofapoptosisatamolecularlevel(DNA-strandbreaks)andidentificationofcellsattheveryearlystagesofapoptosis
EnzymeSolution(TdT),5vials
LabelSolution(fluorescein-dUTP),5vials
