Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Multi-Species

Cited:

Human, Mouse, Rat

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry, ELISA Capture

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG3 Clone # 306507
View all Nitrotyrosine Primary Antibodies »

Product Specifications

Immunogen

Nitrotyrosine-modified KLH

Specificity

Detects Nitrotyrosine adducts on proteins in Western blots. It does not cross-react with phosphotyrosine or 4‑hydroxynonenal adducts. Unfixed cells, tissues, and proteins can be treated with Peroxynitrite (Catalog # AR006) for use as positive controls with this antibody.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG3

Scientific Data Images for Nitrotyrosine Antibody

Detection of Nitrotyrosine antibody by Western Blot.

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 antibody in Human PBMCs by Immunocytochemistry (ICC).

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.

Applications for Nitrotyrosine Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs), A549 human lung carcinoma cells (Positive)immersion fixed HepG2 human hepatocellular carcinoma cells (Positive)

Western Blot

1 µg/mL
Sample: Peroxynitrite or peroxyvanadate-treated NIH‑3T3 mouse embryonic fibroblast cell line

Reviewed Applications

Read 1 review rated 4 using MAB3248 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in TBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Additional Nitrotyrosine Products

Product Documents for Nitrotyrosine Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Nitrotyrosine Antibody

For research use only

Citations for Nitrotyrosine Antibody

Customer Reviews for Nitrotyrosine Antibody (1)

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Customer Images

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  • Nitrotyrosine Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: human cornea
    Species: Human
    Verified Customer | Posted 10/11/2017
    Corneal section stained with anti-nitrotyrosine antibody (1:100) followed by alexa fluor 488.
    Nitrotyrosine Antibody MAB3248

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Protocols

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FAQs

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