Fig. 1.

Overview of the cysteines in Ref-1 and the PEGm redox assay. A) 3D structure of Ref-1 demonstrating the position of the seven cysteine residues in the protein; B–C) A Schematic of the gel-based redox shift assay examining dynamic alteration of Ref-1 redox status in the cells. A reduced Cys residue will be labeled by the PEGm reagent resulting in a 15 kDa shift as shown in B and C. The mixed conformations of Ref-1 protein generate band-shifts based on the protein size and available cysteines and were detected by Ref-1 primary antibody (D).