Illustrated Handbook for High Resolution of IAA Oxidase-Peroxidase Isoenzymes by Isoelectric Focusing in Slabs of Polyacrylamide Gel

摘要

Specific techniques are presented for high resolution of peroxidase-IAA oxidase isoenzymes in polyacrylamide gel slabs by isoelectric focusing in pH gradients. Banding patterns are entirely reproducible. pH gradients are linear and there is no 'cathode shift.' The enzyme stains are very sensitive to low catalytic activity. Since the extreme cathodic or anodic isoenzymes of peroxidase-IAA oxidase enzyme have isoelectric points beyond the limits of wide range carrier ampolytes, it was necessary to adopt a more realistic definition of runtime. Also, a labeling scheme similar to that used in electrophoresis was adapted for isoelectric focusing by use of a standard 'marker isoenzyme.' The number of peroxidase-IAA oxidase isoenzymes revealed by this technique is more than twice that previously reported in the literature.