|Detection of CML-adducted Protein by Western Blot. Western blot shows 100-fold diluted samples of sera from control, Type 1 diabetic, and Type 2 diabetic volunteers. PVDF membrane was probed with 1 µg/mL Carboxymethyl Lysine Monoclonal Antibody (Catalog # MAB3247) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Greater band intensity for the IgG heavy chain (IgG hc) suggests increased glycation in diabetic sera. For additional reference, carboxymethyl lysine-adducted (CML) Leptin and carboxyethyl lysine-adducted (CEL) Leptin were included. A specific band for CML-adducted Leptin was detected (as indicated). This experiment was conducted under reducing conditions using Immunoblot Buffer Group 1.|
N epsilon -carboxymethyl lysine (CML) is formed by the non-enzymatic Schiff-base reaction of glucose with proteins, followed by an Amadori rearrangement and oxidation that leaves only a carboxymethyl group attached to the lysine. The levels of CML adducts accumulate over time and have been used as an indicator of both serum glucose levels and oxidative protein damage. Elevated serum CML-modified proteins have been associated with diabetes and may contribute to diabetic retinopathy, nephropathy and angiopathy.
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