Yakhyayev A V, Voronkova I M, Sukhanov V A
Research Center of Molecular Diagnostics, USSR Ministry of Health, Moscow.
Electrophoresis. 1991 Sep;12(9):680-2.
A method is described for isoelectric focusing of proteins, using an ultrathin-layer polyacrylamide gel on cellophane, followed by electrophoretic transfer of separated proteins onto a nitrocellulose membrane. The polyacrylamide gel is firmly attached to the cellophane and thus protected from mechanical damage; such gels are easily manipulated. Cellophane is permeable to ions and application of this gel support overcomes difficulties resulting from the removal of ultrathin gels from a plastic support on electroblotting. Proteins separated under nondenaturing conditions were transferred onto a nitrocellulose membrane and detected by the concanavalin A-peroxidase technique. The proposed approach makes it possible to analyze the variability of nondenatured proteins and glycoproteins of different origin.
本文描述了一种蛋白质等电聚焦方法，该方法使用玻璃纸上的超薄层聚丙烯酰胺凝胶，然后将分离的蛋白质电泳转移到硝酸纤维素膜上。聚丙烯酰胺凝胶牢固地附着在玻璃纸上，因此可免受机械损伤；这种凝胶易于操作。玻璃纸可透过离子，使用这种凝胶支持物克服了在电印迹时从塑料支持物上取下超薄凝胶所带来的困难。在非变性条件下分离的蛋白质被转移到硝酸纤维素膜上，并通过伴刀豆球蛋白A-过氧化物酶技术进行检测。所提出的方法使得分析不同来源的非变性蛋白质和糖蛋白的变异性成为可能。
