ChIPresultsobtainedwiththeantibodydirectedagainstAML1-ETO
ChIPassayswereperformedusingKasumi-1cells,theantibodyagainstAML1-ETO(Cat.#25208)andoptimizedprimerpairsforqPCR.Shearedchromatinfrom1.25millioncellsand4µlofantibodywereusedperChIPexperiment.QPCRwasperformedusingprimersspecificfortheFUT7,NFE2andOGG1genes.Figure1showstheoccupancy,calculatedastheratio+control/backgroundforwhichthepromoteroftheH2Bgenewasused.

ChIP-seqresultsobtainedwiththeantibodydirectedagainstAML1-ETO
ChIPwasperformedasdescribedabove.TheimmunoprecipitatedDNAof6ChIPassayswaspooledandanalyzedwithanIlluminaGenomeAnalyzer.Librarypreparation,clustergenerationandsequencingwereperformedaccordingtothemanufacturer"sinstructions.The32bptagswerealignedtothehumanreferencegenome(hg18)usingtheELANDalgorithm.Figure2showstheresultsofthecompletechromosome3andthreegenomicregionssurroundingtheOGG1,FUT7andNFE2genes,respectively.ThepositionofthePCRampliconisindicatedwithanarrow.

Determinationoftheantibodytiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstAML1-ETO(Cat.#25208).Theplateswerecoatedwiththepeptidesusedforimmunizationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure3),thetiteroftheantibodywasestimatedtobe1:32,750.

WesternblotanalysisusingtheantibodydirectedagainstAML1-ETO
NuclearextractsofSKNO-1cells(15µg)wereanalyzedbyWesternblotusingtheantibodyagainstAML1-ETO(Cat.#25208)diluted1:1,000inTBS-Tweencontaining5%skimmedmilk.TheMarker(inkDa)isshownontheleft,thepositionoftheproteinofinterestisindicatedontheright. 

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