Agarose native gel electrophoresis of proteins

Abstract

We have developed an agarose-based native gel electrophoresis system that works for both acidic and basic proteins using histidine-MES buffer. This electrophoresis can be done in a flat-bed mode or a vertical mode. While in the flat-bed mode both acidic and basic proteins can be simultaneously analyzed, the vertical gel can only be used for either protein. We have observed that while the migration of acidic bovine serum albumin (BSA) was independent of the buffer concentration, the behavior of basic lysozyme was greatly improved at higher buffer concentration, e.g., 100 mM histidine-100 mM MES. With this buffer system, BSA, lysozyme and chymotrypsin showed expected band mobility and Adeno associated virus particle and bovine gamma globulin showed apparent basic nature of the surface properties.

Keywords: Agarose; Electrophoresis; Flat-bed; Native gel; Proteins; Vertical gel.