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Bpsbioscience/AP1 Reporter Kit (JNK Pathway)/60612/500 reactions

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Background:Thestress-activatedproteinkinase/c-junN-terminalkinase(SAPK/JNK)familyofproteinsincludesmitogen-activatedproteinkinases(MAPKs)thatareactivatedbystress,inflammatorycytokines,mitogens,oncogenes,andinducersofcelldifferentiationandmorphogenesis.UponactivationoftheSAPK/JNKpathway,MAPKinaseKinasesphosphorylateandactivateJNKs.TheactivatedJNKstranslocatetothenucleuswheretheyphosphorylateandactivatetranscriptionfactorssuchasc-Jun.Theactivatedc-Junformshomodimersorheterodimerswithfosfamilyproteinswhichbindtotheactivatorprotein-1(AP1)responseelementandinducetargetgenetranscription.Description:TheAP1ReporterKitisdesignedformonitoringtheactivityoftheJNKsignalingpathwayandthetranscriptionalactivityofAP1inculturedcells.Thekitcontainsatransfection-readyAP1luciferasereportervector.ThisreportercontainsthefireflyluciferasegeneunderthecontrolofmultimerizedAP1responsiveelementslocatedupstreamofaminimalpromoter.TheAP1reporterispremixedwithaconstitutively-expressingRenilla(seapansy)luciferasevectorthatservesasaninternalcontrolfortransfectionefficiency.Thekitalsoincludesanon-inducIBLefireflyluciferasevectorpremixedwithconstitutively-expressingRenillaluciferasevectorasanegativecontrol.Thenon-inducibleluciferasevectorcontainsthefireflyluciferasegeneunderthecontrolofaminimalpromoter,withoutanyadditionalresponseelements.Thenegativecontroliscriticalfordeterminingpathway-specificeffectsandthebackgroundluciferaseactivity.Synonym(s):JNKreporter,SAPKreporterSuppliedAs:Reporter:AP1luciferasereportervector+constitutively-expressingRenillaluciferasevectorNegativeControlReporter:Non-inducibleluciferasevector+constitutively-expressingRenillaluciferasevectorConcentration:BoththeReporterandNegativeControlReporterareprovidedat60ngDNA/µl(500µleach)Format:assaykitInstructionsforuse:Seedatasheetfordetailedtransfectionandassayprotocols.Storage/StABIlity:Stableatleast12monthsfromdateofreceipt,whenstoredasdirected(-20°C)Application(s):•MonitorcAMP/PKAsignalingpathwayactivity.•ScreenforactivatorsorinhibitorsofPKAorcAMP/PKApathwaycomponents.•StudyeffectsofRNAiorgeneoverexpressionontheactivityofthecAMP/PKApathway.Reference(s):1.ZhouH.etal.(2005)FrequencyanddistributionofAP-1sitesinthehumangenome.DNAResearch.11:139-150.2.GaillardP.et.al.(2005)Designandsynthesisofthefirstgenerationofnovelpotent,selective,andinvivoactive(benzothiazol-2-yl)acetonitrileinhibitorsofthec-JunN-terminalkinase.JMedChem.48(14):4596-4607.Notes:Thevectorsaresuitablefortransienttransfection.TheyareNOTsuitablefortransformationandamplificationinbacteria.Warning(s):Avoidfreeze/thawcyclesScientificCategory:CellSignalingPathwayProductType:ReporterAssayKitDatashownislot-specific.Contactusforspecificinformationonotherlots.

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