Mócz G, Bálint M
Anal Biochem. 1984 Dec;143(2):283-92. doi: 10.1016/0003-2697(84)90664-x.
An improved system for polyacrylamide gel electrophoresis in the presence of cationic detergents, cetyltrimethylammonium bromide and cetylpyridinium chloride, respectively, is described. An acidic discontinuous buffer system generated according to the theory of multiphasic zone electrophoresis developed by T. M. Jovin (1973, Biochemistry 12, 871-904) was used. It was optimized with respect to the operational conditions and to the desirable range of relative mobility values for the proteins that have molecular weights from 16,500 to 90,300. Also presented is a procedure for the elimination of interference from cationic detergents frequently encountered during staining of gels. The electrophoretic system was suitable for fractionating a wide variety of proteins. The technique can also be used to provide an alternative estimate of molecular weight. To fully account for accurate estimations, the Ferguson relationship between mobility and gel concentration and the relation of molecular weight to mobility at a single gel concentration were both considered. Examples reported in this paper include the separation and/or molecular weight determination of several common proteins, histones, and microfibrillar and myofibrillar proteins. The results suggest that electrophoresis in the presence of cationic detergents offers the same degree of reliability in analysis of most proteins as is provided by the anionic detergent sodium dodecyl sulfate electrophoresis.
本文描述了一种改进的聚丙烯酰胺凝胶电泳系统，该系统分别在阳离子去污剂十六烷基三甲基溴化铵和十六烷基氯化吡啶存在的情况下运行。使用了根据T. M. Jovin（1973年，《生物化学》12卷，871 - 904页）提出的多相区带电泳理论生成的酸性不连续缓冲系统。针对操作条件以及分子量在16,500至90,300之间的蛋白质的相对迁移率值的理想范围进行了优化。还介绍了一种消除凝胶染色过程中经常遇到的阳离子去污剂干扰的方法。该电泳系统适用于分离多种蛋白质。该技术还可用于提供分子量的另一种估计方法。为了充分考虑准确的估计，同时考虑了迁移率与凝胶浓度之间的弗格森关系以及在单一凝胶浓度下分子量与迁移率的关系。本文报道的例子包括几种常见蛋白质、组蛋白以及微原纤维和肌原纤维蛋白的分离和/或分子量测定。结果表明，在阳离子去污剂存在下进行电泳，在分析大多数蛋白质时，其可靠性与阴离子去污剂十二烷基硫酸钠电泳相当。
