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通过连续的十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳和反相高效液相色谱法分离蛋白质。

Separation of proteins by consecutive sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and reversed phase high performance liquid chromatography.

作者信息

Kaplan B, Pras M

机构信息

Heller Institute of Medical Research, Chaim Sheba Medical Center, Tel-Hashomer, Israel.

出版信息

Biomed Chromatogr. 1991 Mar;5(2):86-9. doi: 10.1002/bmc.1130050209.

Abstract

A procedure for the preparative separation of proteins was developed by using consecutively sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (SDS-PAGE) and reversed phase high performance liquid chromatography (HPLC). The proteins were separated by SDS-PAGE and afterwards extracted from the gel. The extracted proteins were separated from SDS and other small molecular weight contaminants on a Fractogel TSK HW-40 (F) column in acidic aqueous acetonitrile. The proteins eluted from the Fractogel column were fractionated by HPLC. The identity and purity of the recovered proteins was confirmed by SDS-PAGE analysis.

摘要

通过连续使用十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳(SDS-PAGE)和反相高效液相色谱(HPLC),开发了一种蛋白质制备分离方法。蛋白质通过SDS-PAGE进行分离,然后从凝胶中提取。提取的蛋白质在酸性乙腈水溶液中,于Fractogel TSK HW-40(F)柱上与SDS和其他小分子污染物分离。从Fractogel柱上洗脱的蛋白质通过HPLC进行分级分离。回收蛋白质的身份和纯度通过SDS-PAGE分析得以确认。

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