Roche/11684795910 / In Situ Cell Death Detection Kit, Fluorescein现货

• 
  

• 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

1. EnzymeSolution(TdT),5vials

2. LabelSolution(fluorescein-dUTP),5vials

•