Fig. 3.
Effects of diamide and dithiothreitol on IDH. (A) The IDH1/IDH2C56S/C242S mutant enzyme which contains IDH2 Cys-150 as the only cysteine residue was incubated with increasing concentrations of diamide prior to non-denaturing gel electrophoresis (with staining using Coomassie blue) and activity assays. (B) The diamide-treated IDH1/IDH2C56S/C242S mutant enzyme was incubated with increasing concentrations of dithiothreitol (DTT) prior to non-denaturing gel electrophoresis and activity assays. The disulfide bond form of IDH2 migrates aberrantly slowly under these conditions [38, 40], so only portions of the gels are shown.