Production of Monoclonal Antibodies Against Mammalian Ring1B Proteins

Abstract

The Polycomb group (PcG) genes play a role of transcriptional repressor for long-term maintenance of the Hox cluster gene expression. Recently two structurally similar gene products, Ring1A and Ring1B, were identified. Genetic evidence has indicated that Ring1A has Pc-G properties, however, Ring 1B functions are still unknown. To gain functional insights for Ring1B, we raised the mouse monoclonal antibodies (MAbs) against murine Ring1B protein. Using these antibodies, we have detected specifically mouse, human and monkey Ring1B gene products from whole cell extracts in immunoblot and immunoprecipitation analyses. Immunofluorescent staining by the antibodies has shown that endogenous Ring1B proteins clearly co-localize with Ring1A at the pattern of diffuse nuclear speckles. Together with their sequence similarity, Ring1B also may function as a Pc-G protein. Finally, we have proposed that the anti-Ring1B would be useful for biochemical and cytological analyses of Ring1B and Pc-G complex.