hh Nitrotyrosine Antibody MAB3248: R&D Systems

Nitrotyrosine Antibody

(1 citations)   
  • Specificity
    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.
  • Source
    Monoclonal Mouse IgG3 Clone # 306507
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Nitrotyrosine-modified KLH
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunocytochemistry
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; Catalog # NL007) and counterstained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

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 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
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    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.
Background: Nitrotyrosine

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.

  • Alternate Names:
    Nitrotyrosine
Related Research Areas
Citations:

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.

1 Citations: Showing 1 - 1

  1. Age-related retinopathy in NRF2-deficient mice.
    Authors: Zhao Z, Chen Y, Wang J
    PLoS ONE, 2011;6(4):e19456.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Frozen
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

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