Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Sulfotyrosine Monoclonal Antibody, Clone 7C5. Tissue: Embryonic kidney epithelial cell line (HEK293). Species: Human. Fixation: 5% Formaldehyde for 5 min. Primary Antibody: Mouse Anti-Sulfotyrosine 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) DAPI (blue) nuclear stain. (B) Phalloidin Alex Fluor 633 F-Actin stain. (C) Sulfotyrosine Antibody. (D) Composite. Courtesy of: Dr. Robert Burke, University of Victoria.
Western Blot analysis of Sulfotyrosine-BSA Conjugate showing detection of 67 kDa Sulfotyrosine protein using Mouse Anti-Sulfotyrosine Monoclonal Antibody, Clone 7C5. Lane 1: Molecular Weight Ladder (MW). Lane 2: Sulfotyrosine-BSA. Lane 3: Tyrosine-BSA. Lane 4: Phosphotyrosine-BSA. Lane 5: BSA. Load: 2 ug. Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-Sulfotyrosine 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.
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