Starch gel electrophoresis of enzymes—A compilation of recipes

TLDR: Most of the methods outlined here have been used primarily for tissue extracts from mammalian species, and a few have been developed only on plants or lower animals, but this is not to say that they will not work on higher organisms.

Abstract: The technique of starch gel electrophoresis of enzymes with specific staining for activity in the gel, the so-called zymogram method developed by Hunter and Markert (1957), has found many research applications. Screening studies, comparing relatively large numbers of enzymes among a variety of tissues and organisms, are finding increasing use in research on population genetics, taxonomy, etc. The methods presently available are scattered through the literature. This compilation is published in response to numerous requests from investigators in many fields. Most of the methods outlined here have been used primarily for tissue extracts from mammalian species. A few have been developed only on plants or lower animals, but this is not to say that they will not work on higher organisms. The buffer systems have been for the most part empirically arrived at. All are probably subject to improvement. The techniques of making the gels and carrying out the electrophoresis are not described here. These have been amply presented in a number of publications. The vertical system is essentially that described by Shaw and Koen (1968b). The horizontal system is essentially as described by Beckman and Johnson (1964). The horizontal method has the advantage of employing simpler and less expensive apparatus, and it can be run at room temperature using ice trays to keep the gels cold. For many enzyme systems, it provides as good results as the vertical. Both Connaught starch (Connaught Laboratories, Toronto) and Electrostarch (Otto Hiller Company, Madison, Wisconsin) will give satisfactory results with either technique. Unfortunately, different batches of starch may produce varying results, both in rate of migration and in resolution; also, some batches contain substances