Western blot analysis of plasma proteins from two-dimensional gel electrophoresis. Western blot analysis of plasma proteins from two-dimensional gel electrophoresis. Plasma samples from a representative diabetic subject were chosen for performing two-dimensional gel electrophoresis followed by Western blot. Western blot was performed on two-dimensional gel electrophoresis of equal amounts of globulin-depleted plasma (top panels) as well as albumin and globulin-depleted plasma (lower panels) from T2D− (left panels) and T2D+ (right panels) samples. The Western blot showed increased glycation for several plasma proteins from the samples of T2D−. The corresponding gel spots were excised from the Coomassie blue-stained two-dimensional gels of respective samples and analyzed for protein identification by tandem mass spectrometry. All of the identified proteins showed either a reduction or absence of glycation after the insulin treatment (T2D+). The data are from three reproducible experiments done separately on the same samples. Abdul Jaleel et al. Dia Care 2005;28:645-652 ©2005 by American Diabetes Association