Rabilloud Thierry
CEA-DSV-iRTSV/CBM and UMR CNRS-UJF 5249, CEA Grenoble, Grenoble, France.
Methods Mol Biol. 2012;893:25-35. doi: 10.1007/978-1-61779-885-6_2.
Two-dimensional gel electrophoresis has been instrumental in the development of proteomics. Although it is no longer the exclusive scheme used for proteomics, its unique features make it a still highly valuable tool, especially when multiple quantitative comparisons of samples must be made, and even for large samples series. However, quantitative proteomics using 2D gels is critically dependent on the performances of the protein detection methods used after the electrophoretic separations. This chapter therefore examines critically the various detection methods (radioactivity, dyes, fluorescence, and silver) as well as the data analysis issues that must be taken into account when quantitative comparative analysis of 2D gels is performed.
二维凝胶电泳对蛋白质组学的发展起到了重要作用。尽管它已不再是蛋白质组学所使用的唯一方案,但其独特特性使其仍然是一种非常有价值的工具,特别是当必须对样品进行多次定量比较时,甚至对于大量样品系列也是如此。然而,使用二维凝胶的定量蛋白质组学严重依赖于电泳分离后所使用的蛋白质检测方法的性能。因此,本章将严格审视各种检测方法(放射性、染料、荧光和银染)以及在对二维凝胶进行定量比较分析时必须考虑的数据分析问题。