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Signosis/NRF2/ARE Luciferase Reporter NIH3T3 Stable Cell Line/SL-0047-NP/1 Ea
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Description:NRF2/AREResponsiveLuciferaseReporterNIH3T3StableCellLineisderivedfromMousefibroblast,andstablyexpressfireflyluciferasereportergeneunderthecontroloftheNRF2/AREresponseelement. ThiscelllineisanidealcellularmodelformonitoringtheactivationofAntioxidantResponsePathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:NRF2playsacrucialroleincellularanti-oxidantdefense,makingitatherapeutictargetforneurodegenerativediseasesandcancer. Undernormalconditions,NRF2localizesinthecytosolandisrapidlydegradedbytheproteasome. Underoxidativestress,NRF2isstABIlizedandtranslocatestothenucleuswhereitbindstoaDNApromoterandinitiatesgeneexpression. Inthenucleus,NRF2formsaheterodimerwithasmallMafproteinandbindstotheAntioxidantResponseElementintheupstreampromoterregionofmanyantioxidativegenes,andinitiatestheirtranscription.ThisNRF2luciferasereporterstablecelllinehasbeenstablytransfectedwithpTA-ARE-luciferasereportervector,whichcontains4repeatsofantioxidantresponsebindingsites,aminimalpromoterupstreamofthefireflyluciferasecodingregion,alongwitha hygromycinexpressionvector. Followingselection,the hygromycinresistantclonesweresubsequentlyscreenedforTBHQ-inducedluciferaseactivity.Theclonewiththehighestfoldinductionwasselectedandexpandedtoproducethisstablecellline. PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. Data:AnalysisoftheNRF2PathwayReporterNIH3T3StableCellLineinresponsetostimuli. Thecellswereseededona96-wellplatefor8hoursorovernightwithDMEMincluding10%FBS.Thecellsthenweretreatedwithorwithout10μMTBHQor10μM4HNE respectivelyinDMEMand0.1%FBSfor16hours.
Signosis 是一家专业的生物分析供应商,致力于基于微孔板分析技术的生命科学研究产品的开发、生产和销售。Signosis 成立于2007年,坐落于世界研发中心--美国硅谷的中心地带。公司专注于细胞因子、转录因子、miRNA及疾病标志物等重要调控因子和标志物的研究,基于自有专利技术,开发完成400多种先进而独特的用于发现、筛选和分析的生命科学研究产品。Signosis 产品销售遍布北美、欧洲、亚洲及中东等主要国家和地区,客户涵盖美国国立卫生研究院、哈佛大学、耶鲁大学等著名研究机构以及众多制药公司
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