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利用低成本的“自制”中性pH十二烷基硫酸钠-聚丙烯酰胺凝胶电泳系统改进二维凝胶电泳

Improvements in two-dimensional gel electrophoresis by utilizing a low cost "in-house" neutral pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis system.

作者信息

Graham David R M, Garnham Christopher P, Fu Qin, Robbins Jeffrey, Van Eyk Jennifer E

机构信息

Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21224, USA.

出版信息

Proteomics. 2005 Jun;5(9):2309-14. doi: 10.1002/pmic.200401249.

Abstract

Two-dimensional gel electrophoresis (2-DE) is widely used for initial protein separation in proteomics. Commercial products using neutral pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis ((SDS-PAGE)/(Bis (2-hydroxyethyl) imino-tris (hydroxymethyl) methane-HCl, or Bis-Tris)) have greatly improved this technique, but cost and limited sizes restrict their applications. An "in-house" system is presented, resulting in better resolution, separation, and new spot visualization and improved resolution when compared to Tris-HCl gels. Their utility is demonstrated using albumin-depleted serum samples, rabbit heart left ventricle, and human immunodeficiency virus type 1 (HIV-1).

摘要

二维凝胶电泳(2-DE)在蛋白质组学中被广泛用于蛋白质的初步分离。使用中性pH十二烷基硫酸钠-聚丙烯酰胺凝胶电泳((SDS-PAGE)/(双(2-羟乙基)亚氨基-三(羟甲基)甲烷-盐酸,或双三羟甲基氨基甲烷))的商业产品极大地改进了这项技术,但成本和有限的尺寸限制了它们的应用。本文介绍了一种“自制”系统,与Tris-HCl凝胶相比,该系统具有更好的分辨率、分离效果以及新的斑点可视化效果和更高的分辨率。使用去除白蛋白的血清样本、兔心脏左心室和1型人类免疫缺陷病毒(HIV-1)证明了它们的实用性。

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