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使用短毛细管进行蛋白质的十二烷基硫酸钠 - 凝胶电泳。

Sodium dodecyl sulfate-gel electrophoresis of proteins employing short capillaries.

作者信息

Manabe T

机构信息

Department of Chemistry, Faculty of Science, Ehime University, Matsuyama, Japan.

出版信息

Electrophoresis. 1995 Aug;16(8):1468-73. doi: 10.1002/elps.11501601243.

Abstract

The performance of capillary gel-sodium dodecyl sulfate (SDS) electrophoresis for molecular-mass-dependent separation of proteins has been examined employing cross-linked polyacrylamide gels and untreated fused-silica capillaries of short length. An apparatus for capillary electrophoresis has been constructed, combining a UV detector equipped with a capillary holder, a power supply, and a micromanipulator for sample loading. The apparatus is capable of accommodating gel capillaries of different lengths and can be operated with two methods of sample loading: manual injection on top of the gel or electrokinetic injection. Standard proteins have been separated according to their molecular mass (ranging from 17000 to 116000) within 30 min, employing gel capillaries of effective lengths shorter than 10 cm and polyacrylamide gels ranging from 2.4 to 4.8%T (5%C). The results confirmed that gel capillaries of effective length less than 10 cm can be used for protein size-separation by SDS electrophoresis, with much higher performance than is achieved in rod gel-SDS electrophoresis.

摘要

采用短长度的交联聚丙烯酰胺凝胶和未处理的熔融石英毛细管,研究了毛细管凝胶 - 十二烷基硫酸钠(SDS)电泳对蛋白质进行分子量依赖性分离的性能。构建了一种毛细管电泳装置,该装置结合了配备毛细管支架的紫外检测器、电源和用于进样的微操纵器。该装置能够容纳不同长度的凝胶毛细管,并且可以通过两种进样方法进行操作:在凝胶顶部手动进样或电动进样。使用有效长度小于10 cm的凝胶毛细管和2.4%T至4.8%T(5%C)的聚丙烯酰胺凝胶,在30分钟内根据标准蛋白质的分子量(范围为17000至116000)对其进行了分离。结果证实,有效长度小于10 cm的凝胶毛细管可用于通过SDS电泳进行蛋白质大小分离,其性能比棒状凝胶 - SDS电泳要高得多。

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