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Millipore/APT420 | CaspaTag™ Pan-Caspase In Situ Assay Kit, Fluorescein/APT420/25 assays
  • Millipore/APT420 | CaspaTag™ Pan-Caspase In Situ Assay Kit, Fluorescein/APT420/25 assays

Millipore/APT420 | CaspaTag™ Pan-Caspase In Situ Assay Kit, Fluorescein/APT420/25 assays

价格: ¥3048.00 市场价: 5080.00

货号: APT420
品牌: Millipore
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    • Description
      CatalogueNumberAPT420
      BrandFamilyChemicon®
      TradeName
      • CaspaTag
      • Chemicon
      DescriptionCaspaTag™Pan-CaspaseInSituAssayKit,Fluorescein
      BackgroundInformationApoptosisisanevolutionarilyconservedformofcellsuicide,whichfollowsaspecializedcellularprocess.Thecentralcomponentofthisprocessisacascadeofproteolyticenzymescalledcaspases.Theseenzymesparticipateinaseriesofreactionsthataretriggeredinresponsetopro-apoptoticsignalsandresultinthecleavageofproteinsubstrates,causingthedisassemblyofthecell1.

      CaspaseshavebeenidentifiedinorganismsrangingfromC.eleganstohumans.Themammaliancaspasesplaydistinctrolesinapoptosisandinflammation.Inapoptosis,caspasesareresponsIBLeforproteolyticcleavagesthatleadtocelldisassembly(effectorcaspases),andareinvolvedinupstreamregulatoryevents(initiatorcaspases).Anactivecaspaseconsistsoftwolargeandtwosmallsubunitsthatformtwoheterodimers,whichassociateinatetramer2-4.Incommonwithotherproteases,caspasesaresynthesizedasprecursorsthatundergoproteolyticmaturation,eitherautocatalyticallyorinacascadebyenzymeswithsimilarspecificity5.

      Caspaseenzymesspecificallyrecognizea4or5aminoacidsequenceonthetargetsubstrate,whichnecessarilyincludesanasparticacidresidue.Thisresidueisthetargetforcleavage,whichoccursatthecarbonylendoftheasparticacidresidue6.Caspasescanbedetectedviaimmunoprecipitation,immunoblottingtechniquesusingcaspasespecificantibodies,orbyemployingFluorochromesubstrates,whichbecomefluorescentuponcleavagebythecaspase.
      MaterialsRequiredbutNotDelivered·Culturedcellswithmedia

      ·Reagentstoinduceapoptosis

      ·15mLpolystyrenecentrifugetubes

      ·Ambervialsortubesforstorageof150Xconcentrateat-20°C

      ·600mLgraduatedcylinder

      ·Microscopeslides

      ·Hemocytometer

      ·Centrifuge(400xg)

      ·37°Cincubator

      ·Vortexer

      ·Adjustablevolumepipettorwithdisposabletips

      ·Deionizedwater

      ·PBS,pH7.4

      ·DMSO
      ProductInformation
      Components
      • FLICAReagent(FAM-VAD-FMK):Onelyophilizedvial
      • 10XWashBuffer:15mL
      • Fixative:6mL
      • PropidiumIodide:1mLat250μg/mL,ready-to-use
      • HoechstStain:1mLat200μg/mL,ready-to-use
      DetectionmethodFluorescent
      StorageandShippingInformation
      StorageConditions·Storeunopenedkitmaterialsat2-8°Cuptotheirexpirationdate.

      ·ReconstitutedFLICAReagent(150X)shouldbefrozenat-20°Cforupto6months,andmaybethawedtwiceduringthistime.Aliquotintoseparateambertubesifdesired.Protectfromlightatalltimes.

      ·Storediluted(1X)washbufferupto-2-8°Cfor2weeks.
      Applications
      ApplicationTheCaspaTagPan-CaspaseInSituAssayKit,Fluoresceinforflowcytometryisafluorescent-basedassayfordetectionofactivecaspasesincellsundergoingapoptosis.
      KeyApplications
      • FlowCytometry
      • ActivityAssay
      ApplicationNotesTheCHEMICON®CaspaTag™Pan-CaspaseInSituAssayKit,Fluoresceinisafluorescent-basedassayfordetectionofactivecaspasesincellsundergoingapoptosis.Thekitisforresearchuseonly.Notforuseindiagnosticortherapeuticprocedures.

      TestPrinciple

      CHEMICON®"sCaspaTag™InSituCaspaseDetectionKitsuseanovelapproachtodetectactivecaspases.ThemethodologyisbasedonFluorochromeInhibitorsofCaspases(FLICA).Theinhibitorsarecellpermeableandnon-cytotoxic.Onceinsidethecell,theinhibitorbindscovalentlytotheactivecaspase7.Thiskitusesacarboxyfluorescein-labeledfluoromethylketonepeptideinhibitorofcaspase(FAM-VAD-FMK),whichproducesagreenfluorescence.Whenaddedtoapopulationofcells,theFAM-VAD-FMKprobeenterseachcellandcovalentlybindstoareactivecysteineresiduethatresidesonthelargesubunitoftheactivecaspaseheterodimer,therebyinhibitingfurtherenzymaticactivity.Theboundlabeledreagentisretainedwithinthecell,whileanyunboundreagentwilldiffuseoutofthecellandiswashedaway.Thegreenfluorescentsignalisadirectmeasureoftheamountofactivecaspasepresentinthecellatthetimethereagentwasadded.Cellsthatcontaintheboundlabeledreagentcanbeanalyzedby96-wellplate-basedfluorometry,fluorescencemicroscopy,orflowcytometry.
      BIOLOGicalInformation
      SpeciesReactivity
      • Mammals
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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