Establishment of a monoclonal antibody for human LXRalpha: Detection of LXRalpha protein expression in human macrophages.
Establishment of a monoclonal antibody for human LXRalpha: Detection of LXRalpha protein expression in human macrophages.
复制标题
建立人lxralpha的单克隆抗体：在人巨噬细胞中检测lxralpha蛋白的表达。
DOI：
10.1186/1478-1336-1-1
复制
发表时间：
2003-05-09
期刊：
影响因子：
--
通讯作者：
Kodama, Tatsuhiko
中科院分区：
文献类型：
--
作者：
Watanabe, Yuichiro;Tanaka, Toshiya;Uchiyama, Yasutoshi;Takeno, Tetsu;Izumi, Akashi;Yamashita, Hisahiko;Kumakura, Junko;Iwanari, Hiroko;Shu-Ying, Jiang;Naito, Makoto;Mangelsdorf, David J;Hamakubo, Takao;Kodama, Tatsuhiko
肝脏 X 激活受体 α (LXRα) 与 RXR 形成功能性二聚体核受体，调节多种重要脂质的代谢，包括胆固醇和胆汁酸。与RXR相比，细胞内的LXRα蛋白水平较低，并且LXRα蛋白本身很难检测到。我们之前曾报道过 LXRα 的 mRNA 在人类培养的巨噬细胞中高表达。为了证实人巨噬细胞中存在LXRα蛋白，我们建立了针对LXRα的单克隆抗体K-8607。 mAb K-8607 与人 LXRα 蛋白的结合通过多种不同技术得到证实，包括免疫印迹、免疫组织化学和电泳迁移率变动测定 (EMSA)。通过使用该抗体进行免疫印迹，证明了原代培养的人巨噬细胞中存在天然 LXR 蛋白，而人单核细胞中则不存在该蛋白。这种单克隆抗 LXRα 抗体应该被证明是分析人类 LXRα 蛋白的有用工具。
Liver X activated receptor alpha (LXRα) forms a functional dimeric nuclear receptor with RXR that regulates the metabolism of several important lipids, including cholesterol and bile acids. As compared with RXR, the LXRα protein level in the cell is low and the LXRα protein itself is very hard to detect. We have previously reported that the mRNA for LXRα is highly expressed in human cultured macrophages. In order to confirm the presence of the LXRα protein in the human macrophage, we have established a monoclonal antibody against LXRα, K-8607. The binding of mAb K-8607 to the human LXRα protein was confirmed by a wide variety of different techniques, including immunoblotting, immunohistochemistry, and electrophoretic mobility shift assay (EMSA). By immunoblotting with this antibody, the presence of native LXR protein in primary cultured human macrophage was demonstrated, as was its absence in human monocytes. This monoclonal anti-LXRα antibody should prove to be a useful tool in the analysis of the human LXRα protein.