首页 > BPS Bioscience > Bpsbioscience/Notch1/CSL Reporter Kit (Notch Signaling Pathway)/60509/500 reactions

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Bpsbioscience/Notch1/CSL Reporter Kit (Notch Signaling Pathway)/60509/500 reactions

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60509

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4000-520-616

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Background:TheNotchsignalingpathwaycontrolscellfatedecisionsinvertebrateandinvertebratetissues.Notchsignalingistriggeredthroughthebindingofatransmembraneligand,suchasDeltalikeligand4(DLL4)orjagged1(JAG1)toaNotchtransmembranereceptor(Notch1,Notch2,Notch3,Notch4)onaneighboringcell.ThisresultsinproteolyticcleavageoftheNotchreceptor,releasingtheconstitutivelyactiveintracellulardomainofNotch(NICD).NICDtranslocatestothenucleusandassociateswithtranscriptionfactorsCSL(CBF1,RBPJ,SuppressorofHairless,Lag-1)andthecoactivator,Mastermind,toturnontranscriptionofNotchresponsivegenes.Description:TheNotchPathwayReporterkitisdesignedformonitoringtheactivityofNotchsignalingpathwayintheculturedcells.Thekitcontainsthetransfection-readyexpressionvectorforNotch1withtheentireextracellulardomaindeleted(Notch1DE).Insidethecells,theNotch1DEcanbecleavedby-secretaseandactiveNotch1NICDisreleasedtonucleus.ThekitalsocontainsaCSL(CBF1/RBP-J)luciferasereportervector,whichisaNotchpathway-responsivereporter.ThisreportercontainsafireflyluciferasegeneunderthecontrolofmultimerizedCSLresponsiveelementupstreamofaminimalpromoter.TheCSL(CBF1/RBP-J)reporterispremixedwithconstitutivelyexpressingRenilla(seapansy)luciferasevector,whichservesasaninternalpositivecontrolfortransfectionefficiency.Thekitalsoincludesanon-inducIBLefireflyluciferasevectorpremixedwithconstitutivelyexpressingRenillaluciferasevectorasanegativecontrol.Thenon-inducibleluciferasevectorcontainsafireflyluciferasegeneunderthecontrolofaminimalpromoter,butwithoutanyadditionalresponseelements.Thenegativecontroliscriticalfordeterminingpathwayspecificeffectsandbackgroundluciferaseactivity.Synonym(s):Notchreporter,PKAreporter,cAMPreporter,luciferasereporter,CSLreporter,CBF1reporter,RBP-JreporterConcentration:TheReporterandNegativeControlReporterareprovidedat60ngDNA/µl(500µleach).TheNotch1ExpressionVectorandNegativeControlExpressionVectorareprovidedat100ngDNA/µl(250µleach).Format:COMPONENTSInstructionsforuse:Seedatasheetfordetailedtransfectionandassayprotocols.Storage/StABIlity:Stableatleast12monthsfromdateofreceipt,whenstoredasdirected(-20°C)Application(s):•ScreenforactivatorsorinhibitorsoftheNotchsignalingpathway.•StudyeffectsofRNAiorgeneoverexpressionontheactivityoftheNotchpathway.Reference(s):1.LuFMetal.(1996).Proc.Natl.Acad.Sci.USA93(11):5663-5667.2.Kanungoetal(2008).J.NeuRochem.1062236.ApplicationReference:MicroRNA-126-5ppromotesendothelialproliferationandlimitsatherosclerosisbysuppressingDlk1(2014)Notes:Thevectorsaresuitablefortransienttransfection.TheyareNOTsuitablefortransformationandamplificationinbacteria.Warning(s):Avoidfreeze/thawcyclesScientificCategory:CellSignalingPathwayProductType:ReporterAssayKitDatashownislot-specific.Contactusforspecificinformationonotherlots.

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