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单克隆抗体和抗血清识别的分子的分离:固相免疫分离技术。

Isolation of molecules recognized by monoclonal antibodies and antisera: the solid phase immunoisolation technique.

作者信息

Tamura G S, Dailey M O, Gallatin W M, McGrath M S, Weissman I L, Pillemer E A

出版信息

Anal Biochem. 1984 Feb;136(2):458-64. doi: 10.1016/0003-2697(84)90244-6.

Abstract

A simple technique for the isolation of antigens recognized by antisera and monoclonal antibodies has been developed. This method, the solid-phase immunoisolation technique, employs the protein-binding properties of polyvinylchloride microtiter plates. Antibodies are adsorbed to the plates either directly or via an anti-immunoglobulin reagent. Antigen is then placed in the wells, and allowed to adsorb to the antibody. The well is washed, and the antigen is then eluted with a denaturing electrophoresis sample buffer for one- or two-dimensional analysis. The solid-phase immunoisolation technique has been used to isolate a variety of cell membrane antigens with high signals and low backgrounds. The ease of the procedure and the high signal-to-noise ratio make this method preferable to the use of a staphylococcal adsorbent for many applications.

摘要

已开发出一种用于分离抗血清和单克隆抗体所识别抗原的简单技术。这种方法,即固相免疫分离技术,利用了聚氯乙烯微量滴定板的蛋白质结合特性。抗体可直接或通过抗免疫球蛋白试剂吸附到板上。然后将抗原置于孔中,使其吸附到抗体上。洗涤孔后,用变性电泳样品缓冲液洗脱抗原,用于一维或二维分析。固相免疫分离技术已用于分离多种具有高信号和低背景的细胞膜抗原。该方法操作简便且信噪比高,在许多应用中比使用葡萄球菌吸附剂更具优势。

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