将IgG快速印迹到硝酸纤维素膜上，同时使IgM污染最小化。

Rapid blotting of IgG to nitrocellulose with minimal IgM contamination.

作者信息

Hoffman W L, Jump A A

出版信息

J Immunol Methods. 1985 Feb 11;76(2):263-71. doi: 10.1016/0022-1759(85)90303-5.

Abstract

A rapid method is described for the non-electrophoretic transfer of IgG from 0.5 mm thick agarose gels to nitrocellulose. Since the agarose gels are attached to a solid support, the blot is unidirectional. However, 90% of the IgG is transferred in 10 min with no visible loss of resolution. In this procedure, less than 2% of the IgM antibodies are transferred from the gel to the nitrocellulose membrane. Therefore, this technique can be used in IgG spectrotype analysis or antigen-specific assays without the prior removal of IgM antibodies. Approximately 5-6 h are needed to run the gels, blot, and develop the protein pattern with double-antibody immunoperoxidase staining.

摘要

本文描述了一种将IgG从0.5毫米厚的琼脂糖凝胶非电泳转移至硝酸纤维素膜的快速方法。由于琼脂糖凝胶附着于固体支持物上，印迹是单向的。然而，90%的IgG在10分钟内即可转移，分辨率无明显损失。在此过程中，不到2%的IgM抗体从凝胶转移至硝酸纤维素膜。因此，该技术可用于IgG光谱分型分析或抗原特异性检测，无需事先去除IgM抗体。用双抗体免疫过氧化物酶染色进行凝胶电泳、印迹和显影蛋白质图谱大约需要5 - 6小时。