Fig. 7.
Effect of prior reduction and alkylation of disulfide bonds (using TCEP and IAA) on the extent of 1O2-induced glutathionylation of aLA, Lyso and CRP. Panel A: Representative immunoblot image of glutathionylated aLA detected by the anti-GSH antibody for samples without and with prior TCEP/IAA treatment. Panel B: Representative immunoblot image of glutathionylated Lyso detected using an anti-GSH antibody for samples without and with prior TCEP/IAA treatment. Panel C: Representative immunoblot image of oxidized CRP reacted with Bio-GSH and probed using a streptavidin-HRP antibody for samples without and with prior TCEP/IAA treatment. Panel D: OD ratio (OD lane n/OD lane 4, n = 4, 8) of glutathionylated aLA, Lyso and CRP from the immunoblotting results (panels A–C) without (white bars) and with (black bars) prior reduction and alkylation of the disulfide bonds. * Indicates p < 0.05 vs. lane 4 (monomer, panels A–C). Each blot is a representative image from one of three experiments carried out on independent samples. Data in panel D are presented as mean ± SD from three independent experiments.