Peluso R W, Rosenberg G H
Anal Biochem. 1987 May 1;162(2):389-98. doi: 10.1016/0003-2697(87)90409-x.
A method for the reproducible and quantitative electrotransfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to a single sheet of either Zetabind or Gene Screen Plus membranes is presented. This procedure uses commercially available equipment and includes three crucial parameters: the omission of methanol from the transfer buffer, the use of thin (0.75-mm) resolving gels, and a newly developed protocol for pretreatment of the polyacrylamide gel after electrophoresis and before electroblotting. This combination of parameters yields a blot that both qualitatively and quantitatively reflects the proteins in the original polyacrylamide gel.
本文介绍了一种将蛋白质从十二烷基硫酸钠-聚丙烯酰胺凝胶中可重复且定量地电转移至单张Zetabind或Gene Screen Plus膜上的方法。该程序使用市售设备,包括三个关键参数:转移缓冲液中不使用甲醇、使用薄(0.75毫米)的分离胶,以及一种新开发的电泳后和电印迹前聚丙烯酰胺凝胶预处理方案。这些参数的组合产生的印迹在定性和定量上都能反映原始聚丙烯酰胺凝胶中的蛋白质。