Anti peroxiredoxin 3 antibody

MitoSOX Red (3,8-phenanthridinediamine, 5-(6′-triphenylphosphoniumhexyl)-5,6-dihydro-6-phenyl) was purchased from Invitrogen (Invitrogen Life Technologies, CA, USA). Anti-Peroxiredoxin 3 antibody was purchased from Abcam (Abcam, Cambridge, MA, USA). β-Actin Rabbit mAb was purchased from Cell Signaling (Cell Signaling Technology, Inc., MA, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) and TSH were purchased from Sigma (Sigma-Aldrich, MO, USA). RPMI-1640 and FBS were purchased from GE Healthcare Life Sciences (Hyclone, UT, USA). All the other chemicals made in China were of analytical grade.

Rotenone, Antimycin A, goat anti-mouse IgG-HRP, and goat anti-mouse IgG-FITC were bought from Santa Cruz (Santa Cruz Biotechnology, Inc., CA, USA). Anti-Peroxiredoxin-3 antibody was purchased from Abcam (Abcam, Cambridge, MA, USA). Superoxide dismutase (SOD) was bought from Sigma (Sigma-Aldrich, MO, USA). Rat cleaved caspases 3 and 9 ELISA kits were purchased from Chenglin (Chenglin Biotechnology, Beijing, China). TUNEL assay kit was purchased from Boster (Boster, Wuhan, China). FRTL, diethyldithiocarbamate (DETC), MitoSOX Red, MTT, LDH kit, and thyroid stimulating hormone (TSH) were purchased as previously described [2 (link)].

Whole cell proteins were assayed for protein concentration using the bicinchoninic acid (BCA) protein assay kit (Beyotime Institute of Biotechnology, Jiangsu, China). 50 μg of protein was transferred to nitrocellulose membrane followed by SDS polyacrylamide gel electrophoresis. The nitrocellulose membrane was incubated overnight with primary antibodies: Anti-Peroxiredoxin 3 antibody (Abcam, Cambridge, MA, USA) and horseradish peroxidase (HRP) conjugated secondary antibody, developed by Immobilon Western Chemiluminescent HRP Substrate (Merck Millipore, MA, USA). To verify equal loading, β-actin (1 : 1000) (Cell Signaling Technology, Inc., MA, USA) was used as a loading control. Blots were scanned as grayscale images and quantitated using Image J software (NIH). All the blot intensities were normalized with that of loading control β-actin [39 (link)].