Purification of a fragment of MDM2 for production of polyclonal antisera

A histidine-tagged, carboxy-terminal fragment of the murine double minute 2 gene product, p90(MDM2), was purified by Ni-NTA chromatography and prepara tive gel electrophoresis. The purified MDM2 fragment was used to generate p olyclonal antisera that recognize multiple species of MDM2 proteins, includ ing the inhibitor of p53, p90(MDM2), as well as the activator of p53, p76(M DM2). The antibodies are useful for Western analysis, immunoprecipitation, and immunofluorescence. (C) 2001 Academic Press