Nitrotyrosine Antibody

Catalog # Availability Size / Price Qty
MAB3248
MAB3248-SP
Detection of Nitrotyrosine by Western Blot.
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Product Details
Citations (10)
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Nitrotyrosine Antibody Summary

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.
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 either lyophilized or 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
Immersion fixed human peripheral blood mononuclear cells (PBMCs), A549 human lung carcinoma cells (Positive)immersion fixed HepG2 human hepatocellular carcinoma cells (Positive)

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.

Scientific Data

Western Blot Detection of Nitrotyrosine antibody by Western Blot. View Larger

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 Immunoblot Buffer Group.

Immunocytochemistry Nitrotyrosine antibody in Human PBMCs by Immunocytochemistry (ICC). View Larger

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.

Immunocytochemistry View Larger

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.

Immunocytochemistry View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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

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Citations for Nitrotyrosine Antibody

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.

10 Citations: Showing 1 - 10
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  1. Sex-dimorphic moderate hypoglycemia preconditioning effects on Hippocampal CA1 neuron bio-energetic and anti-oxidant function
    Authors: Uddin MM, Ibrahim MMH, Aryal D, Briski KP
    Mol. Cell. Biochem.
  2. Apolipoprotein D modulates amyloid pathology in APP/PS1 Alzheimer's disease mice
    Authors: Hongyun Li, Kalani Ruberu, Sonia Sanz Muñoz, Andrew M. Jenner, Adena Spiro, Hua Zhao et al.
    Neurobiology of Aging
  3. Inflammation Promotes Oxidative and Nitrosative Stress in Chronic Myelogenous Leukemia
    Authors: D ?iki?, A Bogdanovi?, D Markovi?, O Mitrovi?-A, T Suboti?ki, M Dikli?, M Vukoti?, T Dragojevi?, E Živkovi?, JF Santibanez, VP ?oki?
    Biomolecules, 2022-02-03;12(2):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  4. Cellular Response against Oxidative Stress, a Novel Insight into Lupus Nephritis Pathogenesis
    Authors: CD Ene, SR Georgescu, M Tampa, C Matei, CI Mitran, MI Mitran, MN Penescu, I Nicolae
    Journal of personalized medicine, 2021-07-22;11(8):.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Capture
  5. Posttranslational Modifications Pattern in Clear Cell Renal Cell Carcinoma
    Authors: CD Ene, MN Penescu, SR Georgescu, M Tampa, I Nicolae
    Metabolites, 2020-12-27;11(1):.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Capture
  6. Deleterious effects of phosphate on vascular and endothelial function via disruption to the nitric oxide pathway
    Authors: Alan G Jardine
    Nephrol. Dial. Transplant., 2017-10-01;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  7. Age-related retinopathy in NRF2-deficient mice.
    Authors: Zhao Z, Chen Y, Wang J
    PLoS ONE, 2011-04-29;6(4):e19456.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  8. Multi-omic screening of invasive GBM cells in engineered biomaterials and patient biopsies reveals targetable transsulfuration pathway alterations
    Authors: JH Garcia, EA Akins, S Jain, KJ Wolf, J Zhang, N Choudhary, M Lad, P Shukla, S Gill, W Carson, L Carette, A Zheng, S Kumar, MK Aghi
    bioRxiv : the preprint server for biology, 2023-02-24;0(0):.
  9. Nitrosative Stress in the Frontal Cortex From Dogs With Canine Cognitive Dysfunction
    Authors: Prpar Mihevc S, Zako�ek Pipan M, �trbenc M et al.
    Frontiers in Veterinary Science
  10. Phytoestrogen alpha -Zearalanol attenuates homocysteine-induced apoptosis in human umbilical vein endothelial cells
    Authors: Teng Liu, Dan-dan Hou, Qian Zhao, Wei Liu, Pan-pan Zhen, Jian-ping Xu et al.
    BioMed Research International

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