Detects Nitrotyrosine adducts on proteins in Western blots. It does not cross-react with phosphotyrosine or 4‑hydroxynonenal adducts. Cells, tissues, and proteins can be treated with Peroxynitrite (Catalog # AR006) for use as positive controls with this antibody.
Monoclonal Mouse IgG3 Clone # 306507
Protein A or G purified from hybridoma culture supernatant
Lyophilized from a 0.2 μm filtered solution in TBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
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; Catalog # NL007) and counterstained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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 recombinant E. coli DNAK 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 (Catalog # HAF007). The lysates were also probed with Phospho-Tyrosine Monoclonal Antibody (Catalog # MAB1676). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
3-Nitrotyrosine is formed when tyrosine is reacted with peroxynitrite. Since peroxynitrite is formed from nitric oxide and superoxide anion, nitrotyrosine adducts on proteins have been used as markers of oxidative cellular damage and macrophage activation. Elevated nitrotyrosine immunoreactivity has been found in inflammation, osteoarthritis, neurodegenerative diseases, and ischemic damage to the heart and brain.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image. Limited to verified customers in USA, Canada and Europe.