FIGURE 13.

Detection of protein hydroperoxides in RAW 264.7 cell lysates. A, dependence of the concentration of COH formed on the amount of cell lysate added, as expressed by final cellular protein concentration in the reaction mixture. B, dependence of protein hydroperoxides concentration (expressed as COH concentration) in the cell lysates on the time of photolysis. C, effect of ebselen (50 μm) on the concentration of protein hydroperoxides detected in cell lysates by CBA probe. Macrophages RAW 264.7 were preincubated with 5 μm rose bengal, washed with HBSS, and exposed to visible light (100-watt incandescent tungsten light bulb) from 0 to 60 min; next, the cell lysates were prepared and used for further experimentation. The amount of COH produced was normalized to the total amount of protein in the cell lysates. See “Experimental Procedures” for further details. Error bars, S.D.