Single-step, rapid separation of acidic and basic isoenzymes from commercial horseradish peroxidase

A simple and rapid technique for simultaneous separation of the acidic and basic isoenzymes of horseradish peroxidase is described. Upon application of a potential gradient on the electrolyte system composed of Pharmalyte (pH 2.5–5.0) histidine (p I 7.64), and Ampholyte (pH 9–11), the acidic and basic isoenzymes with p I 4.0–8.4 of horseradish peroxidase accumulated at the two interphases generated by this arrangement, with a recovery of 75 ± 5%. Some advantages, such as rapidity, simplicity, low cost, good yields, and others, of this system over existing ones are outlined.