Nichols W L, Gastineau D A, Mann K G
Biochim Biophys Acta. 1979 Jul 5;554(2):293-308. doi: 10.1016/0005-2736(79)90371-7.
High resolution polyacrylamide gel electrophoretic techniques have been applied to the preparative isolation and analysis of plasma membrane proteins and glycoproteins from human platelets and red blood cells. The techniques presented allow relatively simple, direct, rapid and quantitative purification of a broad molecular weight range of membrane proteins, by means of continuous elution preparative gel electrophoresis of protein solubilized with sodium dodecyl sulfate. Spectrophotometric and fluorophotometric (fluorescamine) profiling, and high resolution gel electrophoretic analysis (SDS-acrylamide gradient slab gels, and gel electrofocusing) of eluted protein species indicate that purified membrane proteins of a broad molecular weight range may be obtained in a one step procedure, and in quantities and concentrations sufficient for further analytical or experimental procedures.
高分辨率聚丙烯酰胺凝胶电泳技术已应用于从人血小板和红细胞中制备性分离和分析质膜蛋白及糖蛋白。本文介绍的技术通过用十二烷基硫酸钠溶解的蛋白质进行连续洗脱制备性凝胶电泳,可相对简单、直接、快速且定量地纯化宽分子量范围的膜蛋白。对洗脱的蛋白种类进行分光光度法和荧光光度法(荧光胺)分析以及高分辨率凝胶电泳分析(SDS-丙烯酰胺梯度平板凝胶和凝胶电聚焦)表明,可通过一步法获得宽分子量范围的纯化膜蛋白,其数量和浓度足以用于进一步的分析或实验步骤。