Figure 2. Detection of oxidatively modified cytosolic proteins

TR-iBRB cells were treated with 40 mU/ml glucose oxidase for 4 h. To detect oxidatively modified proteins, we treated the extracts with 2,4 dinitrophenylhydrazine to derivatize protein carbonyls. Equal amounts of derivitized proteins were resolved by SDS-PAGE, and proteins were subsequently transferred to polyvinylidene fluoride (PVDF) membranes and probed with an anti-DNP antibody. The bar graph represents the average (mean±SD) of three independent quantifications of the carbonyls present in protein extracts.