Electrophoretic Analysis of Chemically Modified Taka-amylase A Using a Slab Gel System with Multiple Lanes

Abstract

The limitation of slab gel electrophoresis is in the fixed gel concentration. We devised a slab gel plate to which six different gel concentrations could be applied. Various molecular forms of Taka-amylase A (TAA) prepared by chemical modification were analyzed using this multiple lane system. Modification of Lys, Trp, and carboxyl groups of TAA gave charge isomer forms having different affinities for the substrate, soluble starch. Cross-linked oligomeric forms of TAA were resolved in monomer to tetramer forms in 6.3-12.3% gels. The affinity of TAA for various substrates was also evaluated by the multiple lanes containing different concentrations and various substrates. Moreover, the rapid and simple handling of stained gels by ethanol dehydration and subsequent lapping is described.