Description:
NFkBisaubiquitoustranscriptionfactorthatplaysakeyroleincellularresponsestonumerousstimulisuchasstress,cytokines,ultravioletirrADIation,andbacterialorviralantigens.IncorrectregulationofNFkBhasbeenlinkedtocancer,inflammatoryandautoimmunediseases,andsepticshock.WhenNFkBisactivated,itdissociatesfromitsinhibitorIkBandmovesfromthecytoplasmtothenucleus. Onceinthenucleus,itbindstotargetDNAelementsandpositivelyregulatesthetranscriptionofgenesinvolvedinimmuneandinflammatoryresponses,cellgrowthcontrol,andapoptosis.Principle:
Signosis’proprietaryCDNAplatearrayisaplate-basedhybridizationprofilinganalysisformonitoringtheexpressionofdozensofgenesthroughreversetranscriptionofmRNAintocDNA.Likearrayanalyses,totalRNAisfirstreversetranscribedintocDNAinthepresenceofbiotin-dUTPintheassay.Targetedgenesarethenspecificallycapturedontoindividualwellsonaplate,insteadofmembranes,throughapre-coatedgene-specificoligonucleotide.ThecapturedcDNAsarefurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.Theexpressionlevelofgenesisdirectlyproportionaltotheluminescentintensity. |
Data:
HumanNFkB-regulatedcDNAplatearrayanalysis. HeLacellsweretreatedwith(redbars)orwithout(lightblue)10ng/mlTNFfor30min. Cellswerecollectedforprofiling23geneswiththecDNAplatearrayassay(top).TimecourseofIL-6inductiontoTNFatreatmentinHeLacellswasmonitoredwiththecDNAplatearrayassay.
