Knopf U C, Sommer A, Kenny J, Traut R R
Mol Biol Rep. 1975 Mar;2(1):35-40. doi: 10.1007/BF00357295.
A new method for two-dimensional polyacrylamide gel electrophoresis of proteins is described. The method, illustrated here by its application for the analysis of ribosomal proteins of E. coli, has a high resolving power. The proteins S15 and S16 can be resolved either following alkylation or under reducing conditions. This was not possible with urea gel systems previously employed. The method should be advantageous in the identification of the components of dimers formed with the reagent methyl 4-mercaptobutyrimidate. An additional advantage of the new method is that both dimensions are run at an acidic pH. For ribosomal proteins it is therefore unnecessary to either polymerize the protein sample in the middle of the first dimension disc gel or to electrophorese two samples with opposite polarity.
本文描述了一种蛋白质二维聚丙烯酰胺凝胶电泳的新方法。这里通过其在大肠杆菌核糖体蛋白分析中的应用举例说明该方法具有高分辨率。蛋白质S15和S16在烷基化后或还原条件下均可分离。这在以前使用的尿素凝胶系统中是不可能的。该方法在鉴定与试剂4-巯基丁酸甲酯形成的二聚体成分时应具有优势。新方法的另一个优点是两个维度均在酸性pH下进行。因此，对于核糖体蛋白，无需在第一维圆盘凝胶中间聚合蛋白质样品或电泳两个极性相反的样品。
