Gregory R L, Shechmeister I L
J Immunol Methods. 1983 May 27;60(1-2):141-5. doi: 10.1016/0022-1759(83)90342-3.
A modification of the immunoprecipitation technique was developed to precipitate ribosomal proteins from Streptococcus mutans in isoelectric focusing gels. The method was established in order to determine whether the ribosomal immunogen was contaminated with cell surface proteins. Isoelectric focusing was used to separate the proteins into bands in the gel and various antisera utilized to precipitate the antigens. Electrophoresis of the ribosomal preparation produced 19 protein bands. A rat antiserum raised against the preparation reacted with 8 of these bands and an aliquot of the same antiserum which had been adsorbed with whole S. mutans cells reacted with only 6 bands. This procedure allowed us to detect contamination of the ribosomal preparation with cell surface antigens. The method may be used to detect contamination with cell surface antigens in many different subcellular preparations.
开发了一种免疫沉淀技术的改良方法,用于在等电聚焦凝胶中沉淀变形链球菌的核糖体蛋白。建立该方法是为了确定核糖体免疫原是否被细胞表面蛋白污染。使用等电聚焦将蛋白质在凝胶中分离成条带,并使用各种抗血清沉淀抗原。核糖体制剂的电泳产生了19条蛋白带。针对该制剂产生的大鼠抗血清与其中8条带发生反应,而用完整变形链球菌细胞吸附过的同一抗血清的等分试样仅与6条带发生反应。该程序使我们能够检测核糖体制剂被细胞表面抗原污染的情况。该方法可用于检测许多不同亚细胞制剂中细胞表面抗原的污染。