Schägger H, von Jagow G
Institut für Physikalische Biochemie der Universität München, Federal Republic of Germany.
Anal Biochem. 1987 Nov 1;166(2):368-79. doi: 10.1016/0003-2697(87)90587-2.
A discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) system for the separation of proteins in the range from 1 to 100 kDa is described. Tricine, used as the trailing ion, allows a resolution of small proteins at lower acrylamide concentrations than in glycine-SDS-PAGE systems. A superior resolution of proteins, especially in the range between 5 and 20 kDa, is achieved without the necessity to use urea. Proteins above 30 kDa are already destacked within the sample gel. Thus a smooth passage of these proteins from sample to separating gel is warranted and overloading effects are reduced. This is of special importance when large amounts of protein are to be loaded onto preparative gels. The omission of glycine and urea prevents disturbances which might occur in the course of subsequent amino acid sequencing.
本文描述了一种用于分离1至100 kDa范围内蛋白质的不连续十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）系统。用作尾随离子的三羟甲基氨基甲烷（Tricine），能在比甘氨酸-SDS-PAGE系统更低的丙烯酰胺浓度下实现小蛋白质的分离。无需使用尿素就能实现蛋白质的卓越分离效果，尤其是在5至20 kDa范围内。30 kDa以上的蛋白质在样品凝胶中就已解聚。因此，这些蛋白质能顺利从样品凝胶进入分离凝胶，减少了过载效应。当要在制备凝胶上加载大量蛋白质时，这一点尤为重要。省略甘氨酸和尿素可避免在后续氨基酸测序过程中可能出现的干扰。
