Monoclonal antibodies distinctively recognizing the subtypes of inositol 1,4,5‐trisphosphate receptor: Application to the studies on inflammatory cells

FEBS Letters

Sugiyama, Tomoyasu¹  Yamamoto-Hino, Miki¹  Hasegawa, Mamoru¹  Hanai, Nobuo¹  Monkawa, Toshiaki³  Mikoshiba, Katsuhiko³  Ohmori, Kenji²  Miyawaki, Atsushi³  Furuya, Akiko¹  Satoh, Souichiro² 

[1] Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahimachi, Machida-shi, Tokyo 194, Japan;Pharmaceutical Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 1188 Shimotogari, Nagaizumi-cho, Sunto-gun, Shizuoka-ken 411, Japan;Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108, Japan

关键词: Monoclonal antibody;    Intracellular calcium;    Inositol 1;    4;    5-trisphosphate receptor;    Mast cell;    PMN;    Macrophage;    Thymocyte;    Splenocyte;    Megakaryocyte;    IP3;    inositol 1;    4;    5-trisphosphate;    IP3R;    IP3 receptor;    ELISA;    enzyme linked immunosorbent assay;    PMN;    polymorphonuclear cell;    SD rat;    Sprague—Dawley rat;    FITC;    flourescein isothiocyanate;   

DOI  :  10.1016/0014-5793(94)01099-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Monoclonal antibodies were raised that specifically recognize the COOH-terminal sequences and the loop sequences between the fifth and the sixth transmembrane spanning regions of human inositol 1,4,5-trisphosphate receptor (IP₃R) type 1, 2 and 3. Western blot analysis using Jurkat cells, mouse cerebellum, COS-7 expressing IP₃R type 3 cDNA showed that those monoclonal antibodies reacted specifically with each of these three IP₃R subtypes and that they do not cross-react. These antibodies could be used for the specific immunoprecipitation of IP₃Rs. Using these monoclonal antibodies, the expression profiles of IP₃R-subtype proteins were found to be different among inflammatory cells such as macrophages, polymorphonuclear cells, mast cells, eosinophils, splenocytes, thymocytes and megakaryocytic cells. Usually, more than one type of IP₃R were expressed in a cell simultaneously. The observation of CMK cells under immunofluorescence confocal microscopy revealed that IP₃R type 1 and type 2 are located at different subcellular fractions.

【 授权许可】

Unknown