Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)


  • Product name
    Anti-3-Nitrotyrosine antibody [7A12AF6]
    See all 3-Nitrotyrosine primary antibodies
  • Description
    Mouse monoclonal [7A12AF6] to 3-Nitrotyrosine
  • Host species
  • Specificity
    ab110282 was developed to recognize only protein-bound nitrotyrosine and so is a sensitive tool for measuring protein-specific modifications from oxidative stress.
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, WB, IP, In-Cell ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Nitrated KLH (Keyhole Limpet Hemocyanin).

  • Positive control
    • Flow Cyt: HL-60 cells treated with 2 mM peroxynitrite ICC/IF: HeLa cells and Human fibroblast cells treated with 1 mM peroxynitrite WB: nitrated Bovine heart mitochondria; nitrated tyrosine
  • General notes

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

    Anti-3-Nitrotyrosine antibody (Alexa Fluor® 488) [7A12AF6] (ab157402)

    Anti-3-Nitrotyrosine antibody (HRP) [7A12AF6] (ab198491)

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact or you can find further information here.



Our Abpromise guarantee covers the use of ab110282 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 1 µg/ml.
Flow Cyt Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.


WB Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration.
In-Cell ELISA Use a concentration of 4 µg/ml. (0.4 µg/well).


  • Relevance
    Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling. While multiple nonenzymatic mechanisms are known to be capable of producing nitrated tyrosine residues, most tyrosine nitration events involve catalysis by metalloproteins such as myeloperoxidase, eosinophilperoxidase, myoglobin, the cytochrome P-450s, superoxide dismutase and prostacyclin synthase. Various studies have shown that protein tyrosinenitration is limited to specific proteins and that the process is selective. For example, exposure of human surfactant protein A, SP-A, to oxygen-nitrogen intermediates generated by activated alveolar macrophages resulted in specific nitration of SP-A at tyrosines 164 and 166, while addition of 1.2 mMCO 2 resulted in additional nitration at tyrosine 161. The presence of nitrotyrosine-containing proteins has shown high correlation to disease states such as atherosclerosis, Alzheimer’s disease, Parkinson’s disease and amyotrophic lateral sclerosis.
  • Alternative names
    • Nitrotyrosine antibody


  • Immunocytochemistry image of ab110282 stained Human HeLa cells (A) and fibroblast cells (B, C).
    Cells grown on slides were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15 min). Slides were treated with/without 1 mM peroxynitrite to modify exposed tyrosines to 3-nitrotyrosine. Slides were blocked and incubated with tab110282 at 2 µg/ml overnight at 4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. For reference, the mitochondria (red) were identified by HSP60 / Alexa Fluor® 594 and DAPI was used to stain the cell nuclei (blue).
    HeLa cells (A) and fibroblast cells (B) show surface modification of tyrosine to 3-nitrotyrosine after exposure to peroxynitrite. While (C) unexposed fibroblast cells show no modification.
  • HL-60 cells were stained with 1 µg/ml ab110282 following treatment with 2 mM peroxynitrite (blue) or vehicle control (red). No primary antibody control is shown in black. Peroxynitrite modifies tyrosine residues to 3-nitrotyrosine.
  • All lanes : Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282) at 1 µg/ml

    Lane 1 : Bovine heart mitochondria
    Lane 2 : Bovine heart mitochondria - nitrated
    Lane 3 : BSA
    Lane 4 : BSA - nitrated
  • All lanes : Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282) at 1 µg/ml

    Lane 1 : Bovine heart mitochondria
    Lane 2 : Bovine heart mitochondria - nitrated
    Lane 3 : BSA
    Lane 4 : BSA - nitrated

    Prior to runnning the samples, the membrane was treated with sodium dithionite to reduce nitrotyrosine to aminotyrosine.
  • All lanes : Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282) at 1 µg/ml

    Lane 1 : Bovine heart mitochondria
    Lane 2 : Bovine heart mitochondria - nitrated
    Lane 3 : BSA
    Lane 4 : BSA - nitrated

    In this experiment, the antibody was first blocked with free nitrotyrosine before being used to blot the membrane.
    The figure shows that ab110282's binding capacity was not inhibited by the free nitrotyrosine, and so only binds to the protein-bound form.


This product has been referenced in:
  • Kehm R  et al. Age-related oxidative changes in pancreatic islets are predominantly located in the vascular system. Redox Biol 15:387-393 (2018). Read more (PubMed: 29331666) »
  • Moghimi E  et al. Dietary Vitamin D3 Restriction Exacerbates Disease Pathophysiology in the Spinal Cord of the G93A Mouse Model of Amyotrophic Lateral Sclerosis. PLoS One 10:e0126355 (2015). WB ; Mouse . Read more (PubMed: 26020962) »

See all 6 Publications for this product

Customer reviews and Q&As

Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10% gel)
Guinea pig Tissue lysate - other (cerebellum)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 30 2014

Abcam does not offer peroxynitrite. To generate the datasheet data, we purchased peroxynitrite from Millipore as a 110mM solution in 0.3M NaCl and 0.3M NaOH (Millipore cat # 20-107). The Certificate of Analysis for the product has use instructions, e.g...

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Thank you for contacting us.

Please find the WB protocol used to validate our MitoSciences range of antibodies attached, as well as the pdf datasheets for those products. Keep in mind, we regularly update our online datasheets to reflect the ...

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Thank you for your patience.

I am just following up this case. Would you be so kind to confirm how you are getting on?

Currently, we do not have positive control for this antibody. However, I am very pleased to inform you that after...

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Thank you for getting back to me and for answering to my further questions promptly.

I have been discussing this enquiry with my colleague in the Lab and the methods seem to be chosen all look appropriate ECl, milk block etc, Xray film.

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I am very sorry to hear that both antibodies did not work properly (ab24496 and ab110282).

I can offer you a replacement vial as you wish but it would be important to find out why two antibodies failed to work.

I have read through a...

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Thank you for your enquiry. It is very unfortunate that both of these products (ab24496 and ab110282) do not perform as they are expected to do so.

I would like to reassure you that I take your comment seriously. Though, you have kindly provi...

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Thank you for your enquiry. This antibody is specifically against 3-nitotyorosine only - we are very confident in this. It was generated by nitrated KLH, cross reacts with nitrated BSA (or any other protein) and shows no reaction with un-nitrated KLH....

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Thank you for your response. This is to let you know that I have placed a new order for you - for one vial of ab110282 as a free of charge replacement (exchange for the original item ab24496) and the new order number is 992370. I hope the s...

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Thank you very much for your interest in our anti-Nitrotyrosine antibodies, particularly ab110282. To our knowledge, ab110282 has not been tested in immunohistochemistry on paraffin-embedded sections (IHC-P). Therefore, I can offer a discount off a...

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