Dunn M J, Patel K
Jerry Lewis Muscle Research Centre, Royal Postgraduate Medical School, London, UK.
Methods Mol Biol. 1988;3:217-31. doi: 10.1385/0-89603-126-8:217.
Electrophoretic techniques are, perhaps, the most important group of procedures for the separation and analysis of complex protein mixtures because of their high resolution capacity. This resolving power is maximized in high-resolution, two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) based on the technique developed by O'Farrell (1). This methodology involves a combination of a first-dimension separation in cylindrical isoelectric focusing (IEF) gels containing 9M urea and 2% (w/v) of the detergent NP-40 with discontinuous gradient polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) in the second dimension. Detailed reviews of the theoretical basis, methodological variations, and applications of this technology can be found in refs. 2-5.
由于具有高分辨率能力，电泳技术或许是用于分离和分析复杂蛋白质混合物的最重要的一组方法。基于奥法雷尔（1）开发的技术，这种分辨能力在高分辨率二维聚丙烯酰胺凝胶电泳（2-D PAGE）中得以最大化。该方法包括在含有9M尿素和2%（w/v）去污剂NP-40的圆柱形等电聚焦（IEF）凝胶中进行第一维分离，并在第二维中于十二烷基硫酸钠存在下进行不连续梯度聚丙烯酰胺凝胶电泳（SDS-PAGE）。关于该技术的理论基础、方法变体和应用的详细综述可在参考文献2-5中找到。
