Harper S, Mozdzanowski J, Speicher D
The Wistar Institute, Philadelphia, Pennsylvania, USA.
Curr Protoc Protein Sci. 2001 May;Chapter 10:Unit 10.4. doi: 10.1002/0471140864.ps1004s11.
Two-dimensional gel electrophoresis combines two different electrophoretic separating techniques in perpendicular directions to provide a much greater separation of complex protein mixtures than either of the individual procedures. Variations of the most common two-dimensional technique are described in this unit, namely isoelectrofocusing (IEF) and SDS-PAGE. This unit also includes support protocols describing pI standards and pH profile measurements, casting Immobiline gels, preparation of tissue culture cells and solid tissues for isoelectricfocusing, preparation of molecular weight standards for two-dimensional gels, and two-dimensional protein databases.
二维凝胶电泳将两种不同的电泳分离技术在垂直方向上结合起来,以比任何一种单独的方法更有效地分离复杂的蛋白质混合物。本单元描述了最常见的二维技术的变体,即等电聚焦(IEF)和SDS-PAGE。本单元还包括支持方案,描述了pI标准和pH谱测量、浇铸固定化凝胶、制备用于等电聚焦的组织培养细胞和实体组织、制备二维凝胶的分子量标准以及二维蛋白质数据库。