Shoeman R L, Schweiger H G
J Cell Sci. 1982 Dec;58:23-33. doi: 10.1242/jcs.58.1.23.
Modification of existing two-dimensional techniques enables isoelectric focusing and sodium dodecyl sulphate polyacrylamide gel electrophoresis of complex mixtures of proteins to be completed within 8 h. The method was optimized to separate the protein components of a wheat germ cell-free translation system, providing a statistically proven resolution better than 0 . 03 of a pH unit for the isoelectric point and 1000 for Mr. Fourteen of the more than 300 proteins separated were characterized with respect to Mr and isoelectric point relative to standard proteins under the same conditions. Stained wheat germ proteins thus serve as internal standards for analysis of in vitro translation products.
对现有二维技术的改进使得蛋白质复杂混合物的等电聚焦和十二烷基硫酸钠聚丙烯酰胺凝胶电泳能够在8小时内完成。该方法经过优化，用于分离小麦胚无细胞翻译系统的蛋白质成分，在等电点方面提供了经统计学验证的优于0.03个pH单位的分辨率，在相对分子质量（Mr）方面优于1000。在相同条件下，相对于标准蛋白质，分离出的300多种蛋白质中的14种在相对分子质量和等电点方面得到了表征。因此，染色后的小麦胚蛋白质可作为分析体外翻译产物的内标。
