Isoelectric focusing of proteins separated by SDS-polyacrylamide gel electrophoresis

✍ Scribed by Gen-Ichi Danno

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 779 KB
• Volume: 83
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90525-5

✦ Synopsis

A rapid and simple method for isoelectric focusing of proteins separated by SDS-polyacrylamide gel electrophoresis was described. The focusing system was composed of a polyacrylamide gel layer (7 cm in height) and a solution layer (1.2 cm in height) superimposed on the gel layer. Both layers contained 2% Ampholine, 8 M urea, and 5% sucrose. Protein bands cut into slices from an SDSpolyactylamide gel were washed with 2% Ampholine containing 8 M urea and 5% sucrose, and the washed gel slices were applied directly into the solution layer; then, isoelectric focusing was carried out. Isoelectric points of bovine serum albumin and catalase determined by this method were identical to those of the corresponding protein samples which did not contact with SDS.