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采用非凝胶筛分毛细管电泳法测定丝素蛋白的分子量

Determination of molecular weight of silk fibroin by non-gel sieving capillary electrophoresis.

作者信息

Wei Wei, Zhang Yaopeng, Shao Huili, Hu Xuechao

机构信息

Donghua University, College of Material Science and Engineering, State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, Shanghai 201620, People's Republic of China.

出版信息

J AOAC Int. 2010 Jul-Aug;93(4):1143-7.

Abstract

A simple non-gel sieving capillary electrophoresis (NGSCE) method was established to determine the MW of silk fibroin using CE. The background electrolyte with a pH of 8.8 was based on three components: polyethylene glycol, tris(hydroxymethyl)aminomethane, and sodium dodecyl sulfate (SDS). NGSCE showed a good linear relationship with satisfactory reproducibility between the migration time and the MW of standard proteins. It was found that the regenerated silk fibroin had an MW around 83 kDa with a wide MW distribution (MWD). This absolute value is lower than the result obtained from SDS-polyacrylamide gel electrophoresis due to the different principles of the methods, but their similar MWD shapes indicated that NGSCE could be a feasible, highly sensitive, rapid method for determination of the MW of silk fibroin.

摘要

建立了一种简单的非凝胶筛分毛细管电泳(NGSCE)方法,用于通过毛细管电泳(CE)测定丝素蛋白的分子量。pH值为8.8的背景电解质由三种成分组成:聚乙二醇、三(羟甲基)氨基甲烷和十二烷基硫酸钠(SDS)。NGSCE在迁移时间与标准蛋白质分子量之间显示出良好的线性关系,且重现性令人满意。结果发现,再生丝素蛋白的分子量约为83 kDa,分子量分布(MWD)较宽。由于方法原理不同,该绝对值低于十二烷基硫酸钠-聚丙烯酰胺凝胶电泳得到的结果,但它们相似的MWD形状表明,NGSCE可能是一种可行、高灵敏度、快速测定丝素蛋白分子量的方法。

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