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. Author manuscript; available in PMC: 2009 Oct 28.
Published in final edited form as: Biochemistry. 2008 Oct 2;47(43):11377–11385. doi: 10.1021/bi800952b

Figure 1.

Figure 1

Dimerization of Mtb KatG by reaction with tert-BuOOH in the absence of DMPO and immunochemical detection of protein-derived DMPO nitrone adducts. (A) SDS-PAGE and protein staining. (B) Anti-DMPO immunostain as shown with Western blot. (C) Chemiluminescent detection by ELISA of KatG radical-derived DMPO nitrone adducts. Wild-type KatG (10 μM) exposed to various concentrations of tert-BuOOH (0.01-1 mM) was incubated for 1 h at 37 °C in the presence or absence of 100 mM DMPO in 0.1 M phosphate buffer (pH 7.4). Reactions were stopped by the addition of 20 mM potassium cyanide. Each lane contained 1.6 μg of Mtb KatG. After electrophoresis, the proteins were visualized by being stained with Coomassie blue protein gel stain or transferred to nitrocellulose for Western blot analysis.