Nitrotyrosine Antibody

Detection of Nitrotyrosine by Western Blot. Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 3 mM peroxynitrite, 3 mM inactivated peroxynitrite, or 100 µM peroxyvanadate for 1 hour and recombinantE. coliDNAK treated with 1 mM 4-hydroxynonenal or 1 mM peroxyynitrite for 1 hour. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Nitrotyrosine Monoclonal Antibody (Catalog # MAB3248), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). The lysates were also probed with Phospho-Tyrosine Monoclonal Antibody (MAB1676). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Nitrotyrosine in Human PBMCs. Nitrotyrosine was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using 25 µg/mL Mouse Anti-Nitrotyrosine Monoclonal Antibody (Catalog # MAB3248) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Nitrotyrosine in A549 human lung carcinoma cells Nitrotyrosine was detected in immersion fixed A549 human lung carcinoma cells using Mouse Anti-Nitrotyrosine Monoclonal Antibody (Catalog # MAB3248) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to Cytoplasmic. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Nitrotyrosine in HepG2 cells Nitrotyrosine was detected in immersion fixed HepG2 human hepatocellular carcinoma cells using Mouse Anti-Nitrotyrosine Monoclonal Antibody (Catalog # MAB3248) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to Cytoplasmic. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.