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[一种免疫印迹技术]

[An immunoblot technique].

作者信息

Imai T

机构信息

Faculty of Science, Toho University.

出版信息

Nihon Rinsho. 1995 Sep;53(9):2119-25.

PMID:7474368
Abstract

After a protein is separated by various types of gel electrophoresis, it is transferred onto a nitrocellulose membrane, where a specified protein is detected through immunological reactions. This procedure, called an immunoblot technique, combines gel electrophoresis, which is capable of separating materials and immunological detection with specificity and high sensitivity. It has recently been used widely as a useful method for analyzing proteins in the fields of medicine, biochemistry, molecular biology and human genetics. The aim of this technique is to transfer a target protein, which is present in the gel, as efficiently as possible onto a membrane so that it can react uniquely on the membrane. To achieve this end, it is necessary to select the following; 1) a transfer system, 2) a transfer membrane, 3) a transfer buffer and additive, 4) optimal transfer conditions, and 5) a specific and highly sensitive means of immunological detection, to be combined with the technique. Since the immunoblot technique is, in principle, capable of analyzing all types of proteins as long as there is an antibody specific to the target protein, it can be used widely to determine the physiopathology of human proteins.

摘要

蛋白质经各种类型的凝胶电泳分离后,被转移到硝酸纤维素膜上,在那里通过免疫反应检测特定的蛋白质。这个过程称为免疫印迹技术,它结合了能够分离物质的凝胶电泳以及具有特异性和高灵敏度的免疫检测。最近,它作为一种在医学、生物化学、分子生物学和人类遗传学领域分析蛋白质的有用方法被广泛使用。该技术的目的是将凝胶中存在的目标蛋白质尽可能高效地转移到膜上,以便它能在膜上特异性地发生反应。为了达到这个目的,有必要选择以下各项:1)转移系统,2)转移膜,3)转移缓冲液和添加剂,4)最佳转移条件,以及5)与该技术相结合的特异性和高灵敏度的免疫检测方法。由于免疫印迹技术原则上只要有针对目标蛋白质的抗体就能够分析所有类型的蛋白质,因此它可广泛用于确定人类蛋白质的生理病理学。

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