Simpson Richard J
Cold Spring Harb Protoc. 2010 Apr;2010(4):pdb.prot5412. doi: 10.1101/pdb.prot5412.
Although SDS-PAGE is the method of choice for most denaturing gel electrophoresis procedures, the anionic detergent SDS still presents some drawbacks. For example, SDS forms crystals at low temperatures and, in some cases, causes proteins to aggregate or precipitate. In addition, some proteins are not well-resolved in SDS gels or may migrate anomalously. In these situations, the use of a cationic detergent for PAGE offers an alternative approach. The system described in this protocol uses the cationic detergent cetyltrimethyl ammonium bromide (CTAB) and includes a stacking gel based on the zwitterion arginine (used as a stacking agent) and tricine (N-tris[hydroxymethyl]-methylglycine) used as a counterion and buffer. Some proteins separated on the CTAB electrophoresis system retain their native enzymatic activity, provided the samples are prepared without boiling and without the addition of a reducing agent.
尽管SDS-PAGE是大多数变性凝胶电泳程序的首选方法，但阴离子去污剂SDS仍存在一些缺点。例如，SDS在低温下会形成晶体，并且在某些情况下会导致蛋白质聚集或沉淀。此外，一些蛋白质在SDS凝胶中分辨率不佳或可能异常迁移。在这些情况下，使用阳离子去污剂进行PAGE提供了一种替代方法。本方案中描述的系统使用阳离子去污剂十六烷基三甲基溴化铵（CTAB），并包括一种基于两性离子精氨酸（用作堆积剂）和用作抗衡离子及缓冲剂的三（羟甲基）甲基甘氨酸（tricine）的堆积凝胶。在CTAB电泳系统上分离的一些蛋白质保留了其天然酶活性，前提是样品制备时不煮沸且不添加还原剂。
