Analysis of Protein Glycation Inhibition Using Polyacrylamide Gel Electrophoresis

Nonenzymatic glycation of proteins proceeds over a period, generating harmful advanced glycation end products (AGEs). The accumulation of AGEs is accelerated with chronic hyperglycemia and aging and is implicated in the pathogenesis of chronic diabetic complications and age-related chronic diseases. Assessment of protein glycation requires expensive specialized equipment. The procedure described in this chapter is a simple electrophoresis-based technique that can be used to identify inhibitors of protein glycation. Polyacrylamide gel electrophoresis carried out under native conditions (Native PAGE) separates proteins according to their net charge, shape, and size. As glycation targets free amino groups of proteins, a change in the net charge of the protein occurs following glycation. Since the degree of change in the net charge is proportionate to the degree of glycation, Native PAGE can be used to assess potential inhibitors of protein glycation.