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Establishment of a monoclonal antibody for human LXRα: Detection of LXRα protein expression in human macrophages

Yuichiro Watanabe1,2, Toshiya Tanaka1, Yasutoshi Uchiyama1, Tetsu Takeno2, Akashi Izumi1, Hisahiko Yamashita3, Junko Kumakura3, Hiroko Iwanari3, Jiang Shu-Ying4, Makoto Naito4, David J Mangelsdorf5, Takao Hamakubo1 and Tatsuhiko Kodama1*

Author Affiliations

1 Department of Life Sciences, The University of Tokyo Research Center for Advanced Science and Technology (RCAST), Tokyo, Japan

2 Immunology Research Dept, Tokyo New Drug Research Laboratories II, Pharmaceutical division, Kowa Co, Ltd, Tokyo, Japan

3 Institute of Immunology Co., Ltd, Tokyo, Japan

4 Department of Cellular Function, Division of Cellular and Molecular Pathology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan

5 Department of Pharmacology, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Texas, USA

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Nuclear Receptor 2003, 1:1 doi:10.1186/1478-1336-1-1

Published: 9 May 2003

Abstract

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.