Greaser Marion L, Warren Chad M
University of Wisconsin-Madison, Madison, WI, USA.
Methods Mol Biol. 2012;869:111-8. doi: 10.1007/978-1-61779-821-4_10.
Very large proteins (subunit sizes >200 kDa) are difficult to electrophoretically separate on polyacrylamide gels. A SDS vertical agarose gel system has been developed that has vastly improved resolving power for very large proteins. Proteins with molecular masses between 200 and 4,000 kDa can be clearly separated. Inclusion of a reducing agent in the upper reservoir buffer has been found to be a key technical procedure for obtaining optimum resolution.
非常大的蛋白质(亚基大小>200 kDa)很难在聚丙烯酰胺凝胶上进行电泳分离。已开发出一种SDS垂直琼脂糖凝胶系统,该系统大大提高了对非常大的蛋白质的分辨能力。分子量在200至4000 kDa之间的蛋白质可以清晰分离。已发现在上层储液缓冲液中加入还原剂是获得最佳分辨率的关键技术步骤。