TheMAPK/ERKsignalingpathwayisamajorparticipantintheregulationofcellgrowthanddifferentiation.Itcanbeactivatedbyvariousextracellularstimuliincludingmitogens,growthfactors,andcytokines.Uponstimulation,MEK1/2phosphorylateandactivateERK1/2.TheactivatedERKtranslocatestothenucleuswhereitphosphorylatesandactivatestranscriptionfactors.TheTCFs(TernaryComplexFactors),includingElk1,areamongthebest-characterizedtranscriptionfactorsubstratesofERK.WhenphosphorylatedbyERK,Elk1formsacomplexwithSerumResponseFactor(SRF)andbindstoSerumResponseElement(SRE),resultingintheexpressionofnumerousmitogen-inducIBLegenes.
TheSREReporterKitTransientPackisdesignedtoprovidethetoolsnecessaryfortransientlytransfectingandmonitoringtheactivityoftheERKsignalingpathwayandthetranscriptionalactivityofSREinculturedHEK293cells.Thekitcontainstransfection-readyvectorscontainingfireflyluciferaseasaMAPK/ERKpathway-responsivereporterandconstitutivelyexpressingRenillaluciferaseasatransfectioncontrol. ItalsoincludestheDualLuciferasedetectionreagentstodetectbothluciferaseactivitiesandspecializedmediumforgrowingandassayingHEK293cells.
ThekeytotheSRETransientPackistheSREluciferasereportervector,whichisanERKpathway-responsivereporter.ThisreportercontainsthefireflyluciferasegeneunderthecontrolofmultimerizedSREresponsiveelementslocatedupstreamofaminimalpromoter.TheSREreporterispremixedwithaconstitutively-expressingRenillaluciferasevectorthatservesasaninternalcontrolfortransfectionefficiency.
Thepackalsoincludesanon-induciblefireflyluciferasevectorpremixedwithconstitutively-expressingRenillaluciferasevectorasanegativecontrol.Thenon-inducibleluciferasevectorcontainsthefireflyluciferasegeneunderthecontrolofaminimalpromoter,withoutanyadditionalresponseelements.Thenegativecontroliscriticalfordeterminingpathway-specificeffectsandthebackgroundluciferaseactivity.
Additionally,thepackincludescellculturemedium(BPSMedium1)thathasbeenoptimizedforusewithHEK293cells*.BPSMedium1includesMEMmedium,10%fetalbovineserum,1%non-essentialaminoacids,sodiumpyruvate,and1%Pen/Strep. Finally,thepackprovidestheDualLuciferase(Firefly-Renilla)AssaySystem.Theseluciferasereagentsprovidehighlysensitive,stabledetectionoffireflyluciferaseactivityandRenillaluciferaseactivity. Thedualluciferasereagentscanbeuseddirectlyincellsingrowthmedium,andcanbedetectedwithanyluminometer;automatedinjectorsarenotrequired.
*Othermammaliancelllinesmayalsobeused,butanalternatecellculturemediummayberequiredforoptimalcellgrowth.
•ScreenforactivatorsorinhibitorsoftheMAPK/ERKsignalingpathway.
•StudyeffectsofRNAiorgeneoverexpressionontheactivityoftheMAPK/ERKpathway.
2.Treisman,R.(1992)Theserumresponseelement.TrendsBiochemSci.17(10):423-426.
BPS bioscience总部坐落于世界最大之一的美国圣地亚哥生命科学研究机构和公司集中地的中心。公司主要提供激酶、磷酸二酯酶、磷酸酶、组蛋白去甲基化酶、组蛋白甲基化转移酶、HDACs、聚ADP核糖聚合酶。BPS公司为药物研发用重组酶类的提供了最大选择之一。产品类型 重组酶类；分子细胞生物学；重组蛋白；分析试剂盒；底物；重组细胞系
本网站将在规定时间内给予删除等相关处理。
