Polyacrylamide Gel Electrophoresis after Isoelectric Focusing of Proteins

Abstract

This chapter describes the polyacrylamide gel electrophoresis after isoelectric focusing of proteins. The technique of isoelectric focusing in polyacrylamide gel cylinders is relatively simple and allows several samples to be investigated simultaneously. The chapter presents two-dimensional procedure in which isoelectric focusing is followed by electrophoresis. This two-stage procedure not only allows the smaller ampholyte molecules to migrate ahead of the proteins but also produces a recognizable map which is more easily interpreted. The chapter illustrates a typical example of a protein pattern obtained with normal serum. The gel cylinder in which isoelectric focusing is performed is shown at the top of the illustration and proteins with low isoelectric points are shown to the left, those with high isoelectric points to the right