Porzio M A, Pearson A M
Biochim Biophys Acta. 1977 Jan 25;490(1):27-34. doi: 10.1016/0005-2795(77)90102-7.
Standard experimental procedures for continuous polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate were modified to give more effective separation and improved resolution of myofibrillar proteins. The system utilizes a running gel consisting of 10% acrylamide with 0.1% bisacrylamide crosslinker (100:1) incorporating 400 mM Tris/glycine (pH 8.80), 0.1 mM ethylenediaminetetraacetate, 5% glycerol and 0.1% sodium dodecyl sulfate. Electrophoresis was performed at 1 mA per gel with corresponding running times of 4-6 h. The myosin heavy chain enters and migrates as a narrow symmetrical band while the smaller regulatory proteins of the myofibril are resolved. The utility of the procedure in relation to the study of protein structure is detailed.
对十二烷基硫酸钠存在下连续聚丙烯酰胺凝胶电泳的标准实验程序进行了改进,以更有效地分离肌原纤维蛋白并提高分辨率。该系统使用一种运行凝胶,其由10%丙烯酰胺与0.1%双丙烯酰胺交联剂(100:1)组成,包含400 mM Tris/甘氨酸(pH 8.80)、0.1 mM乙二胺四乙酸、5%甘油和0.1%十二烷基硫酸钠。电泳以每块凝胶1 mA的电流进行,相应的运行时间为4 - 6小时。肌球蛋白重链以狭窄的对称带形式进入并迁移,而肌原纤维较小的调节蛋白则得到分离。详细阐述了该程序在蛋白质结构研究方面的实用性。