MULTIPLE IMMUNOREPLICA TECHNIQUE - SCREENING FOR SPECIFIC PROTEINS WITH A SERIES OF DIFFERENT ANTIBODIES USING ONE POLYACRYLAMIDE-GEL
MULTIPLE IMMUNOREPLICA TECHNIQUE - SCREENING FOR SPECIFIC PROTEINS WITH A SERIES OF DIFFERENT ANTIBODIES USING ONE POLYACRYLAMIDE-GEL
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DOI：
10.1016/0003-2697(81)90577-7
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发表时间：
1981-01-01
影响因子：
2.9
通讯作者：
VERMA, DPS
中科院分区：
文献类型：
--
作者：
LEGOCKI, RP;VERMA, DPS
提出了一种简单的电泳蛋白免疫鉴定方法。一种聚丙烯酰胺凝胶中的蛋白质可以经过一系列的电泳转移到硝化纤维素纸上（部分western-blots），从而得到该凝胶的多个副本。每个副本可以与一系列不同的抗血清（至少3）反应，其中前一个抗体通过pH 2.2处理去除。抗原-抗体复合物使用125I-Protein a可视化。固定在硝化纤维素纸上的蛋白质的反应性和特异性抗原不受反复孵育和低pH处理的影响。抗体检测到的相同大小的复制品和重叠的蛋白质图谱允许在原始凝胶中精确定位特定多肽。
A simple method for immunological identification of proteins resolved electrophoretically is presented. Proteins from 1 polyacrylamide gel can be subjected to a series of electrophoretic transfers to nitrocellulose paper (partial western-blots), providing several replicas of the gel. Each replica can be reacted with a series of different antisera (at least 3), where the preceding antibody is removed by treatment with pH 2.2. The antigen-antibody complexes are visualized using 125I-Protein A. Reactivity and antigenic specifically of proteins immobilized on nitrocellulose paper is not affected by repeated incubations and low pH treatments. Identical size of the replicas and superimposable profiles of proteins detected by antibodies allow a precise localization of particular polypeptides in the original gel.