Werner W E, Demorest D M, Stevens J, Wiktorowicz J E
Applied Biosystems Division of Perkin-Elmer, Foster City, California 94404.
Anal Biochem. 1993 Jul;212(1):253-8. doi: 10.1006/abio.1993.1319.
The determination of molecular weights by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is one of the most powerful electrophoretic techniques for protein characterization. A separation media has been developed which allows this type of analysis to be performed in the capillary format. A replaceable polymeric solution rather than a polymerized gel was used as the sieving matrix. This solution allowed for the separation of proteins denatured in SDS in a size-dependent manner as demonstrated by the linear correlation between the proteins' relative migration and the concentration of the sieving matrix (Ferguson plot). The logarithm of the molecular weight of protein standards correlated linearly with the relative mobility of the denatured proteins over the molecular weight range of 14,000 to 205,000. The calculated resolution at half-peak height was such that proteins that differed by as little as 4% in molecular weight would be resolved. Finally, the integrated peak areas at 215 nm were linearly proportional to the mass of the protein injected.
通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）测定分子量是蛋白质表征最强大的电泳技术之一。已开发出一种分离介质，可使这种类型的分析以毛细管形式进行。使用可替换的聚合物溶液而非聚合凝胶作为筛分基质。该溶液能够以大小依赖的方式分离在SDS中变性的蛋白质，蛋白质的相对迁移与筛分基质浓度之间的线性相关性（弗格森图）证明了这一点。蛋白质标准品分子量的对数与变性蛋白质在14,000至205,000分子量范围内的相对迁移率呈线性相关。计算得出的半峰高处的分辨率使得分子量相差仅4%的蛋白质也能被分离。最后，215 nm处的积分峰面积与注入蛋白质的质量呈线性比例关系。
