Vanfleteren J R
Laboratorium voor Morfologie en Systematiek der Dieren, Rijksuniversiteit Gent, Belgium.
Anal Biochem. 1989 Mar;177(2):388-91. doi: 10.1016/0003-2697(89)90071-7.
A method is described which combines the resolving power of two-dimensional gel electrophoresis with that of acetic acid/urea/Triton X-100 gel electrophoresis, avoiding the necessity of eluting protein from the gels at any step of the procedure. The combination of electrophoretic separation on the basis of charge, mass, and hydrophobic properties of the proteins has the potential of resolving modified forms and isoforms present in very complex protein populations. The technique can be used for analytical purposes, or it may be scaled up to yield microgram amounts of highly purified proteins. The resolution obtained by tandem application of nonequilibrium pH gradient electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and polyacrylamide gel electrophoresis in the presence of nonionic detergent was evaluated using crude nuclear proteins of the nematode Caenorhabditis elegans.
本文描述了一种方法，该方法将二维凝胶电泳的分辨能力与乙酸/尿素/ Triton X - 100凝胶电泳的分辨能力相结合，避免了在该程序的任何步骤中从凝胶上洗脱蛋白质的必要性。基于蛋白质的电荷、质量和疏水特性进行电泳分离的组合，有潜力分辨非常复杂蛋白质群体中存在的修饰形式和同工型。该技术可用于分析目的，也可扩大规模以产生微克量的高度纯化蛋白质。使用线虫秀丽隐杆线虫的粗核蛋白评估了非平衡pH梯度电泳、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和在非离子去污剂存在下的聚丙烯酰胺凝胶电泳串联应用所获得的分辨率。
