蛋白组分抽提试剂盒说明书

OverviewDescriptionReferencesProductInformationApplicationsBiologicalInformationSafetyInformationProductUsageStatementsStorageandShippingInformationSupplementalInformationPrix&DisponibilitéPrix&DisponibilitéRéférenceDisponibilitéConditionnementQtéPrixQuantité539790-1KITEnstockContacterleServiceClients1kitÀlavalidationdelacommandePlusd'informations—AjouterauxfavorisAjouteraupanierDescriptionDescriptionOverviewFastandreproducibleextractionofsubcellularproteomesfrommammaliancellsProteoExtract®SubcellularProteomeExtractionKit(S-PEK)isdesignedforfastandreproducibleextractionofsubcellularproteomesfromadherentandsuspension-grownmammaliancells.TheS-PEKtakesadvantageofthedifferentsolubilitiesofcertainsubcellularcompartmentsinthefourselectedreagents.Inthecaseofadherentcells,theprocedureisperformeddirectlyinthetissueculturedishwithouttheneedforcellremoval.Cellsorthepartsofthecellsremainattachedtotheplateduringsequentialextractionofsubcellularcompartments,untiltheappropriateextractionreagentisused.Thus,theearlydestructionofthecellularstructurebyenzymaticormechanicaldetachmentofcellsfromthetissuecultureplateandanymixingofdifferentsubcellularcompartmentsisprevented.Forsuspension-growncells,extractionstartswithgentlesedimentationandwashingofthecells.Thestepwiseextractiondeliversfourdistinctproteinfractionsfromonesample:Cytosolicfraction(F1)Membrane/organelleproteinfraction(F2)Nucleicproteinfraction(F3)Cytoskeletalfraction(F4)ProteinsareobtainedinthenativestatemakingtheS-PEKsuitableformanydownstreamapplicationssuchas1Dand2Dgelelectrophoresis,immunoblotting,enzymeactivityassays,andproteinmicroarrays.Samplesize:3-5x106or25-50mgtissue.CatalogueNumber539790BrandFamilyCalbiochem®SynonymsS-PEKKitFeaturesandbenefitsStepwiseextractionresultinginfourdistinctsubcellularproteomesfromonesampleHighlyreproducibleNoultracentrifugationstepsFast—needsjust2hourswith45minuteshands-ontimeProducesproteinssuitableforfunctionalstudiesDescriptionApplicationDataA:AdherentSAOScellswereextractedaccordingtotheDetailedProtocolforSubcellularExtractionofProteinsFromAdherentCellsasoutlinedabove.Imagesi-ivshowthemorphologyofthecellsbeforeandaftereachextractionstep(200-foldenlarged).TheSAOScellsremainedattachedthroughouttheextractionprocedure.B:AnaliquotofeachfractionfromAweresubjectedtoSDS-PAGEanalysis(F1-F4=fractions1-4).Thedatademonstratecleardifferencesintheproteinbandingpatternsamongthe4fractions.C:AliquotsofeachfractionfromAwereseparatedbySDS-PAGEandtransferredtoPVDmembraneforblottingwiththeindicatedantibodies.Forc-Fos,thefractionsweresubjectedtoimmunoprecipitation,priortoWesternblotting,toenrichforanyc-Fospresentineachfraction.Thedatademonstratethateachmarkerproteinisspecificallyenrichedwithintheappropriatefraction.DescriptionA431cellswerestimulatedwith0.2µg/mlTNF-αfortheindicatedtimes.AttheendofeachinductionperiodthecellswereextractedasoutlinedintheDetailedProtocolforSubcellularExtractionofProteinsfromAdherentCells.TheproteinsfromanaliquotofeachfractionwereseparatedbySDS-PAGEandtransferredtoPVDmembraneforWesternblotanalysisusinganantibodyspecificforNF-κB.ThedataindicatethatthereismeasurabletranslocationofNF-κBfromthecytoplasmtothenucleasasearlyas5minafterTNF-αstimulation.Description*Testedonratliverandbovinelivertissue.DescriptionDescription