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Signosis/NFkB Luciferase Reporter HepG2 Stable Cell Line/SL-0017-FP/1 Ea

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SL-0017-FP

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4000-520-616

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Description:NFkBResponsiveLuciferaseReporterHepG2StableCellLineisderivedfromhumanlivercancer,andstablyexpressfireflyluciferasereportergeneunderthecontroloftheNFkB responseelement. Thiscelllineisanidealcellularmodelformonitoringtheactivation ofNFkBReceptorSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:NFkBplaysanimportantroleincontrollingmanyBIOLOGicalprocessesincludingimmuneandinflammatoryresponses,developmentalprocesses,cellulargrowth,andapoptosis.Inresponsetothevariousstimuli,suchasstress,cytokines,freerADIcals,ultravioletirradiation,andbacterialorviralantigens,NFkBisactivatedandtranslocatesfromcytoplasmtonucleus,whereNFkBbindstoitsresponseelementonthepromoterregionandregulatesawidespectrumofgeneexpression.DysfunctionofNFkBactivityisassociatedwithcancer,inflammatoryandautoimmunedisease,andviralinfection.MonitoringtheNFkBactivityisessentialtounveilthemechanismofthesediseasesandconductdrugdiscovery.SignosishasestablishedaNFkBluciferasereporterstablecelllinethathasbeenstablytransfectedwithpTA-NFkB-luciferasereportervectorandcanbeusedforstudyingNFkBsignalingpathwaysactivatedbydifferentcytokines,suchasTNFαandmanyotherstimuli,enforcedgeneexpressionandgeneknockdown.ThecelllinewasestablishedbytransfectionusingapTA-NFkB-luciferasereportervector,whichcontains4repeatsofNFkBbindingsites,aminimalpromoterupstreamofthefireflyluciferasecodingregion,alongwithhygromycinexpressionvectorfollowedbyhygromycinselection.ThehygromycinresistantclonesweresubsequentlyscreenedforTNFa-inducedluciferaseactivity. PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. Data: AnalysisofSL-0017NFκBreporteractivityinresponsetoTNFαtreatment. HepG2cellswereseededona96-wellplateforovernightwithDMEMincluding10%FBS.Thecellsthenweretreatedwithorwithout20ng/mlTNFαinDMEMand0.1%FBSfor16hours. HepG2-NFkBLuciferaseReporterCellLineexhibitsTNFα-dependentincreaseinluciferaseactivitywhencomparedtountreatedcells.

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