Rademacher B E, Steele W J
Department of Pharmacology, College of Medicine, University of Iowa, Iowa City 52242.
Anal Biochem. 1987 Nov 15;167(1):37-46. doi: 10.1016/0003-2697(87)90131-x.
Modifications of ISO-DALT devices that further enhance the efficiency and reproducibility of two-dimensional mapping of proteins are described. The principal changes in ISO system devices include the introduction of a gel casting trough with a removable panel to permit the removal of excess gel without introducing air into the electrofocusing gels and the introduction of an upper electrode compartment with a separate watertight septum for each electrofocusing tube to permit tube removal for cleaning and replacement. The principal changes in DALT system devices include the use of modified powder funnels to introduce acrylamide solutions into the slab gel gradient former without aeration; the introduction of a flexible outlet system for the gradient former to facilitate the removal of air bubbles; the introduction of an inexpensive two-part mixing chamber to permit disassembly for cleaning; the use of split gel holders to eliminate deformation and breakage of electrofocusing gels during loading onto slab gels; the introduction of an inexpensive integrated slab gel casting/rotating apparatus; and the introduction of a simple, water-cooled slab gel electrophoresis apparatus to reduce the volume of running buffer used in electrophoresis.
本文描述了对ISO-DALT装置的改进，这些改进进一步提高了蛋白质二维图谱分析的效率和可重复性。ISO系统装置的主要变化包括：引入带有可移除面板的凝胶浇铸槽，以便在不向等电聚焦凝胶中引入空气的情况下去除多余凝胶；引入上电极室，每个等电聚焦管都有一个单独的防水隔膜，以便移除管子进行清洁和更换。DALT系统装置的主要变化包括：使用改进的粉末漏斗在不曝气的情况下将丙烯酰胺溶液引入平板凝胶梯度形成器；为梯度形成器引入一个灵活的出口系统，便于去除气泡；引入一个便宜的两部分混合室，以便拆卸进行清洁；使用分体凝胶支架，以消除等电聚焦凝胶加载到平板凝胶过程中的变形和破损；引入一个便宜的集成平板凝胶浇铸/旋转装置；以及引入一个简单的水冷平板凝胶电泳装置，以减少电泳中使用的运行缓冲液体积。
