Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Dityrosine Monoclonal Antibody, Clone 10A6 . Tissue: Embryonic kidney epithelial cell line (HEK293). Species: Human. Fixation: 5% Formaldehyde for 5 min. Primary Antibody: Mouse Anti-Dityrosine Monoclonal Antibody at 1:50 for 30-60 min at RT. Secondary Antibody: Goat Anti-Mouse Alexa Fluor 488 at 1:1500 for 30-60 min at RT. Counterstain: Phalloidin Alexa Fluor 633 F-Actin stain; DAPI (blue) nuclear stain at 1:250, 1:50000 for 30-60 min at RT. Localization: Cytoplasmic. Magnification: 20X (2X Zoom). (A,C,E,G) - Untreated. (B,D,F,H) - Cells cultured overnight with 50 µM H2O2. (A,B) DAPI (blue) nuclear stain. (C,D) Phalloidin Alexa Fluor 633 F-Actin stain. (E,F) Dityrosine Antibody. (G,H) Composite. Courtesy of: Dr. Robert Burke, University of Victoria.

Western Blot analysis of Dityrosine-BSA Conjugate showing detection of 67 kDa Dityrosine protein using Mouse Anti-Dityrosine Monoclonal Antibody, Clone 10A6 . Lane 1: Molecular Weight Ladder (MW). Lane 2: BSA. Lane 3: 3,5-Dibromotyrosine-BSA. Lane 4: Dityrosine-BSA. Lane 5: Bromotyrosine-BSA. Lane 6: 7-ketocholesterol-BSA. Load: 1 µg. Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-Dityrosine Monoclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:2000 for 60 min at RT. Color Development: ECL solution for 5 min in RT. Predicted/Observed Size: 67 kDa.

Western Blot analysis of Human Cervical cancer cell line (HeLa) lysate showing detection of Dityrosine protein using Mouse Anti-Dityrosine Monoclonal Antibody, Clone 10A6 . Lane 1: Molecular Weight Ladder (MW). Lane 2: HeLa cell lysate. Lane 3: H2O2 treated HeLa cell lysate. Load: 12 µg. Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-Dityrosine Monoclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:2000 for 60 min at RT. Color Development: ECL solution for 5 min in RT.