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2 protocols using rabbit polyclonal anti prdx1

1

Oxidative Stress Protein and DNA Assays

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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.
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2

Oxidative Stress Biochemical Assay

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Tert-butyl hydroperoxide (t-BHP), sodium dodecyl sulfate (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, mouse monoclonal anti-PRDX6, mouse monoclonal anti-4-Hydroxynonenal (4-HNE), rabbit polyclonal anti-catalase, and 8-hydroxy-deoxyguanosine (8-OHdG). Anti-thioredoxin 1 antibody was purchased from Cell Singaling Thecnology (Danvers, MA, USA), Polyvinylidene fluoride (PVDF) membranes (0.22 µm pore size; Osmonics Inc., Minnetonka, 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 (Denville Scientific, Inc., Saint-Laurent, QC, Canada) were also used for immunodetection of blotted proteins. Other chemicals were used at the reagent level.
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