Electrophoretic methods for the isolation and characterization of enzymes Review Article

Electrophoretic methods are effective tools in enzyme isolation, purification and characterization. For preparative purposes mainly polyacrylamide gel electrophoresis, isoelectric focusing and capillary electrophoresis are applied, usually as a final purification step after previous chromatographic separations. More often, electrophoretic methods are utilized for enzyme characterization, that is for molecular weight determination, determination of isoelectric point and for appraisal of enzyme purity. Determination of molecular weight with SDS-PAGE and with non-denaturing method makes possible to calculate the number of subunits in the molecule and their character. Isoelectric point is estimated with isoelectric focusing. For purity evaluation any electrophoretic method is convenient. For detection of enzyme zones on the gel after electrophoresis, specific activity staining or immunoblotting is accessible.