BioVyonProteinA,analternativesolid-phaseaffinitymatrixforchromatinimmunoprecipitationIgorChernukhina,1,SungYunKanga,1,SamBrownb,SvetlanaGrettona,ClaudiaFabiolaMendez-Catalaa,DaveCowiesonb,ElenaKlenovaa,⇑aDepartmentofBiologicalSciences,UniversityofEssex,Colchester,EssexCO43SQ,UKbPorvairTechnology,ADivisionofPorvairFiltrationGroup,WrexhamLL139XS,UKarticleinfoArticlehistory:Received12November2010Receivedinrevisedform25January2011Accepted25January2011Availableonline1February2011Keywords:Chromatinimmunoprecipitation(ChIP)BioVyonProteinAProteinASepharoseDynabeadsProteinACCCTC-bindingfactor(CTCF)CTCFbinding(target)sites(CTSs)RNApolymeraseIITATAboxQuantitativereal-timepolymerasechainreaction(Q-PCR)abstractChromatinimmunoprecipitation(ChIP)isanimportanttechniqueinthestudyofDNA/proteininterac-tions.TheChIPprocedure,however,haslimitationsinthatitislengthy,canbeinconsistent,andispronetononspecificbindingofDNAandproteinstothebead-basedsolid-phasematricesthatareoftenusedfortheimmunoprecipitationstep.Inthisinvestigation,weexaminedtheutilityofanewmatrixforChIPassays,BioVyonProteinA,asolidsupportbasedonporouspolyethylene.InChIPexperimentscarriedoutusingtwoantibodiesandsevenDNAloci,theperformanceofBioVyonProteinAwassignificantlybet-ter,withagreaterpercentageofDNApull-downinalloftheassaystestedcomparedwithbead-basedmatrices,ProteinASepharose,andDynabeadsProteinA.Furthermore,therigidporousdiscformatwithinacolumnmadetheBioVyonmatrixmucheasiertousewithfewerstepsandlessequipmentrequire-ments,resultinginasignificantreductioninthetimetakentoprocesstheChIPsamples.Insummary,BioVyonProteinAprovidesacolumn-basedassaymethodforChIPandotherimmunoprecipitation-basedprocedures;therigidporousstructureofBioVyonenablesafastandrobustprotocolwithhigherChIPenrichmentratios.CrownCopyright�2011PublishedbyElsevierInc.Allrightsreserved.Thechromatinimmunoprecipitation(ChIP)2assayisanimpor-tantresearchtoolinmodernmolecularbiology[1–5].ItallowsthestudyandidentificationofDNAsequencesthatarespecificallybound...
