Description:

NFkBResponsiveLuciferaseReporterMDA-MB-231CellLineisderivedfrom Humanbreastcancer,andstablyexpressfireflyluciferasereportergeneunderthecontrolofNFkBresponseelement.Thiscelllineisanidealcellularmodelformonitoringtheactivationof NFkBresponsePathwaytriggeredbystimulitreatment,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-0043NFκBreporteractivityinresponsetoTNFαandIL-1atreatment. TheMDA-MB-231cellswereseededona96-wellplateforovernightincompletegrowthmedia. Thecellsthenweretreatedwithorwithout20ng/mlTNFαand20ng/mlIL-1aingrowthmediawith0.1%FBSfor16hours. MDA-MB-231NFkBLuciferaseReporterCellLineexhibitsTNFαandIL-1a-dependentincreaseinluciferaseactivitywhencomparedtountreatedcells.

Signosis基因表达  是利用来自基因的信息指导功能性基因产物合成的过程。这些产品通常是蛋白质。基因表达分析对于理解基因及其蛋白产物的功能，以及对影响基本生物学过程的复杂调控网络的清晰了解至关重要。Signosis  提供了许多用于基因表达研究的测定产品，包括用于监控由TF上调或下调的基因表达的cDNA板阵列，以及用于分析基因表达的单细胞PCR试剂盒，可直接分析细胞无需RNA分离即可进行裂解。 SARS-CoV-2实时PCR检测试剂盒单细胞基因表达分析印迹分析RNA的cDNA板阵列细胞裂解液的cDNA板阵列