Production of monoclonal antibodies against mammalian Ring1B proteins
Production of monoclonal antibodies against mammalian Ring1B proteins
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DOI：
10.1089/027245701300060427
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发表时间：
2001-02-01
期刊：
影响因子：
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通讯作者：
Koseki, H
中科院分区：
文献类型：
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作者：
Atsuta, T;Fujimura, S;Koseki, H
Polycomb group（PcG）基因在Hox簇基因表达的长期维持中起着转录抑制因子的作用。最近，两个结构相似的基因产物，Ring 1A和Ring 1B，被确定。遗传证据表明Ring 1A具有Pc-G特性，然而Ring 1B的功能仍然未知。为了获得Ring 1B的功能见解，我们提出了针对小鼠Ring 1B蛋白的小鼠单克隆抗体（MAbs）。用这些抗体，我们已经检测到特异性小鼠，人和猴Ring 1B基因产物的全细胞提取物中的免疫印迹和免疫沉淀分析，免疫荧光染色的抗体已表明，内源性Ring 1B蛋白明确地与Ring 1A共定位在模式的弥漫核斑点。由于Ring 1B与Pc-G蛋白的序列相似性，Ring 1B也可能具有Pc-G蛋白的功能。最后，我们提出了Ring 1B的抗Ring 1B抗体将用于Ring 1B和Pc-G复合物的生化和细胞学分析。
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.