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抗人C1q:制备单特异性抗血清的快速简便方法。

Anti-human C1q: rapid and simple method for preparing monospecific antisera.

作者信息

Yonemasu K

出版信息

J Immunol Methods. 1975 Dec;9(2):185-94. doi: 10.1016/0022-1759(75)90109-x.

Abstract

A new simple, rapid and economical method is described for producing monospecific antisera to human C1q without using gel filtration or column chromatography. Moderately purified C1q is obtained by dialyzing fresh human serum in the presence of chelating agents at low ionic strength and then electrophoresing it in agarose. When injected into rabbits, the electrophoretically purified product induced potent antisera to three or four serum proteins including C1q, all with slow electrophoretic mobilities (gamma to beta) at pH 8.6. The antibodies to serum proteins other than C1q are easily removed by immunoadsorbents consisting of the insolubilized supernatants obtained from the dialysis used to make the original C1q-rich fraction. The monospecific antisera prepared by this technique form only one band of precipitate in the slow gamma region in immunoelectrophoresis with whole human serum or C1q-rich solution, as well as in Ouchterlony double diffusion test. They agglutinate EAClq but not EA cells and detect the same antigen as standard monospecific antisera to human C1q obtained by another well-established method.

摘要

本文描述了一种新的简单、快速且经济的方法,用于制备针对人C1q的单特异性抗血清,无需使用凝胶过滤或柱色谱法。通过在低离子强度下于螯合剂存在的条件下透析新鲜人血清,然后在琼脂糖中进行电泳,可获得中度纯化的C1q。当将这种经电泳纯化的产物注射到兔子体内时,会诱导产生针对包括C1q在内的三或四种血清蛋白的强效抗血清,这些蛋白在pH 8.6时均具有缓慢的电泳迁移率(γ至β)。针对C1q以外血清蛋白的抗体可通过由用于制备原始富含C1q组分的透析所得不溶性上清液组成的免疫吸附剂轻松去除。用该技术制备的单特异性抗血清在与全人血清或富含C1q溶液的免疫电泳中,以及在双向琼脂扩散试验中,仅在缓慢的γ区域形成一条沉淀带。它们能凝集EAC1q但不能凝集EA细胞,并且检测到的抗原与通过另一种成熟方法获得的人C1q标准单特异性抗血清相同。

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