Pollard J W
MRC Human Genetic Disease Research Group, Department of Biochemistry, Queen Elizabeth College, University of London, Campden Hill, London, England.
Methods Mol Biol. 1984;1:81-96. doi: 10.1385/0-89603-062-8:81.
Since O'Farrell (1) introduced the improved technique for high resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), it has become one of the most powerful tools for the separation and quantification of proteins from complex mixtures. The principal reason for this is that the method employs separation of denatured proteins according to two different parameters, molecular weight and isoelectric point. Consequently, it has sufficient resolution to separate individual proteins as discrete spots on the gel. Each parameter may also be varied and therefore, with the modification of non-equilibrium pH-gradient electrophoresis (NEPHGE) to analyze basic proteins (2), almost any polypeptide may be investigated. Thus to date, the O'Farrell 2-D gel system has no serious rivals, with the possible exception of the Kaltschmidt and Wittmann (3) gel system for analyzing ribosomal proteins. Ribosomal proteins, however, may be adequately separated with NEPHGE.
自从奥法雷尔（1）引入了改进的高分辨率二维聚丙烯酰胺凝胶电泳（2-D PAGE）技术以来，它已成为从复杂混合物中分离和定量蛋白质的最强大工具之一。其主要原因是该方法根据分子量和等电点这两个不同参数对变性蛋白质进行分离。因此，它具有足够的分辨率，能够在凝胶上以离散斑点的形式分离单个蛋白质。每个参数也可以变化，因此，通过对非平衡pH梯度电泳（NEPHGE）进行改进以分析碱性蛋白质（2），几乎任何多肽都可以进行研究。因此，迄今为止，奥法雷尔二维凝胶系统几乎没有竞争对手，分析核糖体蛋白质的卡尔施密特和维特曼（3）凝胶系统可能是个例外。然而，核糖体蛋白质可以通过NEPHGE得到充分分离。
