ChIPresultsobtainedwiththeantibodydirectedagainstRARA
ChIPassayswereperformedusingNB4cells,theantibodyagainstRARA(Cat.#25310)andoptimizedprimerpairsforqPCR.Shearedchromatinfrom6millioncellsand4µlofantibodywereusedperChIPexperiment.QPCRwasperformedusingprimersspecificfortheTGM2,HMHA1,PRAM1andH2Bgenes.Figure1showstherelativeoccupancy,calculatedastheratio+control/backgroundforwhichthesecondexonoftheMBgenewasused.

ChIP-seqresultsobtainedwiththeantibodydirectedagainstRARA
ChIPwasperformedasdescribedaboveandtheimmunoprecipitatedDNAwasanalyzedwithanIlluminaGenomeAnalyzer.Librarypreparation,clustergenerationandsequencingwereperformedaccordingtothemanufacturer"sinstructions.The32bptagswerealignedtothehumanreferencegenome(hg18)usingtheELANDalgorithm.Figure2showstheresultsofthecompletechromosome19andtwo50kbregionsurroundingtheHMHA1andPRAM1genes,respectively.

Determinationoftheantibodytiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainsthumanRARA(Cat.#25310).Theplateswerecoatedwiththepeptidesusedforimmunizationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure3),thetiteroftheantibodywasestimatedtobe1:2,400.

WesternblotanalysisusingtheantibodydirectedagainstRARA
Humanembryonickidneycells(293T)weretransfectedwithaRARAconstruct(lane2)orwithanegativecontrolconstruct(lane1)andanalyzedbyWesternblotusingtheantibodyagainstRARA(Cat.#25310),diluted1:750inBSA/PBS-Tween.ThemolecularweightMarker(inkDa)isshownontheleft;thelocationoftheproteinofinterestisindicatedontheright. 

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