Description:

NFkBisaubiquitoustranscriptionfactorthatplaysakeyroleincellularresponsestostimulisuchasstress,cytokines,freerADIcals,ultravioletirradiation,oxidizedLDL,andbacterialorviralantigens.IncorrectregulationofNFkBhasbeenlinkedtocancer,inflammatoryandautoimmunediseases,septicshock,viralinfection,andimproperimmunedevelopment.WhenNFkBisactivated,itisdissociatedfromitsinhibitorIkBandmovesfromthecytoplasmtothenucleus,whereitbindstotargetDNAelementsandpositivelyregulatesthetranscriptionofgenesinvolvedinimmuneandinflammatoryresponses,cellgrowthcontrol,andapoptosis.TogetabetteroverviewofgeneexpressionofNFkBregulatedgenes,SignosishasdevelopedaCDNAPlateArraywhichcouplesitsCL^TM cDNAsynthesiskitwhichallowsreversetranscriptiondirectlyfromcelllysateswithoutRNApreparation.Thisplate-basedassaysallowsprofilingtheexpressionof20+NFkB-regulatedgenesdirectlyincelllysates.Thisassayprovidesquantitativedata,allowingresearcherstoscreenmultiplesamples,aswellashighsensitivity,allowingresearcherstouselesssample. 

Principle:

UnlikeconventionalreversetranscriptionwhichneedstotalRNA,SignosishasdevelopedaCL(TM)cDNAsynthesis-basedplatearray.ItcansynthesizecDNAprobesthroughreversetranscriptiondirectlyincelllysateswithoutRNApreparation.ThesynthesizedcDNAprobesarethenappliedforaplatearrayhybridization.Targetedgenesarespecificallycapturedontoindividualwellsonaplatethroughapre-coatedgene-specificoligonucleotide.ThecapturedcDNAsarefurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.Theexpressionlevelofgenesisdirectlyproportionaltotheluminescentintensity.

 

 

Data:

HumanNFkB-regulatedcDNAplatearrayanalysis. HeLacellsweretreatedwith(redbars)orwithout(lightblue)10ng/mlTNFfor30min. Cellswerecollectedforprofiling23geneswiththecDNAplatearrayassay(top).TimecourseofIL-6inductiontoTNFatreatmentinHeLacellswasmonitoredwiththecDNAplatearrayassay.

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