Nitrotyrosine Antibody
R&D Systems, part of Bio-Techne | Catalog # MAB3248
Conjugate
Catalog #
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
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 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.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)
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
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.
*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.
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
Product Specific Notices for Nitrotyrosine Antibody
For research use only
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