Signosis/ER Filter Plate Assay/FA0003/1 Ea

Description:

ERs(ERalphaandERbeta)mediatecellularsignalingofestrogens,belongingtothenuclearreceptorfamilyoftranscriptionfactors.ThemechanismofregulatoryactionsofERsisthatligand-boundERsbinddirectlytoestrogenresponseelements(ERE),inthepromotersoftargetgenesorviaprotein-proteininteractionswithothertranscriptionfactors.ThedysfunctionofERshasbeendemonstratedtobecloselyrelatedwithbreastcancer,prostatecancerandlungcancer.SignosishasdevelopedERfilterassayformonitoringDNAbindingactivityofER.Unlikeconventionalgelshiftassays,ERfilterassayisahighthroughputassayanditcananalyzemultiplesamplessimultaneously.Meanwhile,theassaycanbesetupforafewsampleswithlesstediousthannon-radioactivegelshiftassayormuchsafethan ³²Pbasedgelshiftassay.

Principle:

InSignosisTFfilterplateassays,aspecificbiotin-labeledTFDNAbindingsequenceismixedwithnuclearextractstoallowformationofTF-DNAcomplexes.AfilterplateisusedtoretainboundDNAprobeandremovefreeprobe.Theboundpre-labeledDNAprobeisthenelutedfromthefilterandcollectedforquantitativeanalysisthroughDNAplatehybridization.ThecapturedDNAprobeisfurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.

Data: