Fig. 5.
Effect of increasing HSA-to-heme ratios on the protein-radical formation in human plasma by heme/H2O2. (A) Coomassie blue-stained gel to show equal loading, (B) anti-DMPO blotting, and (C) anti-DMPO ELISA. Reactions included 30 μM H2O2, 100 mM DMPO, 30 μM heme, and different concentrations of human plasma, expressed here as HSA in plasma (1.5, 7.5, 30, 60, 90 μM). Reaction mixtures were incubated for 2 h at 37 °C in Chelex-treated, 100 mM phosphate buffer, pH 7.4, containing 25 μM DTPA. Data show representative gel/blot or mean values ±SE from at least three experiments in duplicate (n=3).