S-phase detection with an antibody to bromodeoxyuridine. Role of DNase pretreatment

摘要: We have previously described a monoclonal antibody (BU-1) to 5-bromo-2-deoxyuridine (BrdUrd) that is useful for measurement of cell cycle S-phase. BU-1 hybridoma supernatant reacted with incorporated BrdUrd after the cells had been ethanol fixed; without requirement acid or base denaturation. found this reactivity lost if purified used, culture supernatants are heated, mycoplasma-free line isolated. The contained endogenous DNase activity was result mycoplasma infection line. This required staining in absence other denaturation steps. could be substituted by addition bovine pancreatic I. disruption double stranded DNA structure an enzyme rather than harsh chemical heat treatments does not affect protein cellular morphology and allows detection morphologic antigenic subsets. pre-treatment may also 'hidden' antigens.