MonoclonalAntibodytoHumanTopoisomeraseIDescription
ThismonoclonalantibodywasraisedtohumantopoI(TG2005H-RC1)usingstandardmethods. Theantibody(IgGclass)worksverywellforWesternblottingexperiments. Theepitopehasnotbeenmapped. ThisproducthasnotbeentestedforIPorIFapplications;however,giventhehighspecificityseenonWesterns,itshouldworkwellintheseapplications.

TG2012-4contains250unitsofmonoclonalantibodyfromaconcentratedhybridomasupernatant.UnitdefinitionisbasedonWesternblottingunits:oneunitcorrespondstoa1:1000dilutionofantibodysolutionsufficienttodilutedworkingprobe.Thevialcontains250ulofantibodyat1unit/ul.Forexample,toprepare10mlofdilutedantibodyprobe,at10ulofthestockantibodysolutionto10mlofsuitablediluent(TBSforexample).ThiswillbesufficienttodetecthumantopoisomeraseIin10ugofacrudenuclearextractoftypicalcells(HeLa).Notethattheunitconcentrationisveryconservativeandinmanysituations,higherdilutionsoftheproductmayworkwell.Iftheextractsarepreparedfromnuclei,proteolysisislessproblematic.Proceduresformakingnuclearextractsaredescribedhere.Notethatinmostcasesastandardwholecellextractwillalsoworkifusedfreshextractsareprepared.Amethodformakingfreshnuclearextractsisdescribedhere.

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