ChIPresultsobtainedwiththeantibodydirectedagainstMBD1
ChIPassayswereperformedusinghumanosteosarcoma(U2OS)cells,theantibodyagainstMBD1(Cat.#25303)andoptimizedPCRprimersets.Shearedchromatinfrom1x10⁶cellsand1.5µgofantibodywereusedperChIPexperiment.BeadsonlywereusedasanegativeIPcontrol.QuantitativePCRwasperformedwithprimersforthepromotersoftheMLH1gene(usedasapositivecontrol)andCDC6gene(usedasanegativecontrol).Figure1showstherecovery,expressedasa%ofinput(therelativeamountofimmunoprecipitatedDNAcomparedtoinputDNAafterqPCRanalysis).

Determinationoftheantibodytiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainsthumanMBD1(Cat.#25303),crudeserumandflow-through.Theplateswerecoatedwiththepeptideusedforimmunizationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure2),thetiterofthepurifiedantibodywasestimatedtobe1:20,000.

WesternblotanalysisusingtheantibodydirectedagainstMBD1
NuclearextractsofHeLacells(40µg)wereanalyzedbyWesternblotusingtheantibodyagainstMBD1(Cat.#25303)diluted1:500inTBS-Tweencontaining5%skimmedmilk.Thepositionoftheproteinofinterestisindicatedontheright,theMarker(inkDa)isshownontheleft. 

Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.