Bicinchoninic protein determination assay

tert-butyl hydroperoxide (tert-BHP), SDS, phosphotungstic acid, buthylated hydroxytoluene, 2-thiobarbituric acid and malonaldehyde bis(dimethyl acetal), the Bicinchoninic protein determination assay and the anti-α-tubulin were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). The following were purchased from Abcam Inc., (Cambridge, MA, USA): rabbit polyclonal anti-PRDX1, monoclonal anti-PRDX4, monoclonal anti-PRDX6, the antigenic peptide used to raise the anti-PRDX1 antibody and 8-hydroxy-deoxyguanosine (8-OHdG). The anti-8-OHdG antibody was purchased from StressMarq Biosciences Inc., (Victoria, BC, Canada). Biotinylated horse anti-mouse antibody and Horse Serum were purchased from Vector Labs. Alexa-555 fluor streptavidin (1 mg ml⁻¹ in H₂O) and ProLong Gold antifade with DAPI were purchased from Invitrogen Life Technologies (Burlington, ON, Canada). Nitrocellulose (0.22 μm pore size; Osmonics Inc., MN, USA), donkey anti-rabbit IgG and goat anti-mouse IgG, both conjugated to horseradish peroxidase (Cedarlane Laboratories Ltd., Hornby, ON, Canada), an enhanced chemiluminescence kit (Lumi-Light; Roche Molecular Biochemicals, Laval, QC, Canada) and radiographic films (Fuji, Minamiashigara, Japan) were also used for immunodetection of blotted proteins. Other chemicals used were of at least reagent grade.