Quandt N, Stindl A, Keller U
Institut für Biochemie und Molekulare Biologie, Technische Universität Berlin, Federal Republic of Germany.
Anal Biochem. 1993 Nov 1;214(2):490-4. doi: 10.1006/abio.1993.1527.
A convenient method was established for the M(r) estimation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of large polypeptides in the range from 250,000 to 600,000. Such polypeptides had previously been shown to migrate improperly in various gel electrophoresis systems. In a conventional Laemmli system with 3% polyacrylamide, approximately linear log M(r) vs Rf plots could be obtained by varying the gel thickness which ranged from 0.4 to 1.5 mm. In the additional presence of an electrolyte, such as NaCl, running times were shorter and for evaluation linear plots could be constructed from runs in very thin gels with less than 0.5 mm thickness. Handling of gels was greatly facilitated by fixing them to one of the gel chamber glass plates via bind-silane treatment prior to casting.
建立了一种简便的方法，用于通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对分子量在250,000至600,000范围内的大型多肽进行分子量（M(r)）估计。此前已证明此类多肽在各种凝胶电泳系统中迁移异常。在含有3%聚丙烯酰胺的传统Laemmli系统中，通过改变凝胶厚度（范围为0.4至1.5毫米），可以获得近似线性的log M(r)与相对迁移率（Rf）的关系图。在额外存在电解质（如氯化钠）的情况下，运行时间更短，并且为了进行评估，可以从厚度小于0.5毫米的非常薄的凝胶运行中构建线性图。在灌胶前通过结合硅烷处理将凝胶固定到凝胶室玻璃板之一上，极大地方便了凝胶的处理。
