Khalkhali-Ellis Z
Department of Biology, Washington University, St. Louis, MO 63130, USA.
Prep Biochem. 1995 Feb-May;25(1-2):1-9. doi: 10.1080/10826069508010103.
Resolution of a wide range of polypeptides, 3.5-200kDa, on a single low acrylamide and cross linkage gel of 7.7% T, 2.6% C is described here. Laemmli's (4) original discontinuous SDS polyacrylamide gel electrophoresis (SDS-PAGE) system is modified by increasing the ionic strength of both stacking and resolving gels, and replacing the usual glycine buffer with a tricine cathode buffer as described by Schagger and von Jagow (7). This system offers the advantage of a wide range of protein fractionation, with sufficient band resolution, on a single, low acrylamide concentration and cross linkage gel. Moreover, increased gel ionic concentration allows higher protein and salt load, and renders this system suitable for preparative work.
本文描述了在一块单一的低丙烯酰胺和交联度(7.7% T,2.6% C)凝胶上对3.5 - 200kDa的多种多肽进行分离的方法。Laemmli(4)最初的不连续SDS聚丙烯酰胺凝胶电泳(SDS - PAGE)系统经过了改良,增加了堆积胶和分离胶的离子强度,并按照Schagger和von Jagow(7)的描述,用三羟甲基氨基甲烷阴极缓冲液替代了常用的甘氨酸缓冲液。该系统具有在单一低丙烯酰胺浓度和交联度凝胶上实现广泛蛋白质分级分离的优势,且具有足够的条带分辨率。此外,增加的凝胶离子浓度允许更高的蛋白质和盐负载量,使该系统适用于制备工作。