• Product name
    Nitrotyrosine ELISA Kit
  • Detection method
  • Assay type
  • Assay duration
    Multiple steps standard assay
  • Product overview

    This nitrotyrosine (3-nitrotyrosine, 3NT) competitive ELISA is used to determine the level of nitrotyrosine modified proteins in a sample. The provided microplates have been evenly coated with a nitrotyrosine containing antigen. The competitive ELISA is performed by adding the test sample mixed with the provided HRP conjugated anti-3NT antibody. If no 3NT modified proteins are present in the sample, all of the anti-3NT detector antibody is available to bind to the immobilized 3NT containing protein coating the wells, generating a maximum (100%) binding signal. In contrast, if soluble 3NT modified protein is present in the sample, it will compete for binding by the anti-3NT detector antibody. The degree of competition is proportional to the concentration of soluble 3NT modified proteins in the sample. Therefore the signal in each well has an inverse relationship to the amount of 3NT in each sample. When performing each assay a standard curve is generated from the provided standard to allow the accurate quantitation of the 3NT content in the test samples.

    The assay is followed by monitoring the HRP dependant color change in each well at 600 nm. Alternatively, the assay can be terminated, at a user-defined time, by the addition of 1N HCl (not supplied) and the assay performed as an end-point measurement at 450 nm

  • Tested applications
    Suitable for: Competitive ELISAmore details
  • Platform



Our Abpromise guarantee covers the use of ab113848 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
Competitive ELISA Use at an assay dependent concentration.


  • Example standard curve in 3NT BSA equivalents.

  • The same example standard curve in 3NT molarity.



This product has been referenced in:
  • Magnifico MC  et al. Nonylphenol and Octylphenol Differently Affect Cell Redox Balance by Modulating the Nitric Oxide Signaling. Oxid Med Cell Longev 2018:1684827 (2018). Read more (PubMed: 29805728) »
  • Biswal MR  et al. Timing of Antioxidant Gene Therapy: Implications for Treating Dry AMD. Invest Ophthalmol Vis Sci 58:1237-1245 (2017). Read more (PubMed: 28241311) »

See all 9 Publications for this product

Customer reviews and Q&As

I am pleased to let you know that there is a separate extraction buffer available which is the same as the one provided for ab113848 Nitrotyrosine ELISA Kit:

ab156035  Native lysis Buffer
(25mM Hepes, 100mM NaCl,1.5% Lauryl Maltos...

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As explained in the protocol page 17 (https://www.abcam.com/ps/products/113/ab113848/documents/ab113848%20Nitrotyrosine%20ELISA%20Kit_24%20June%202014_JC%20(website).pdf) the final concentration and molarity of the standard dilution series takes into a...

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This ELISA is a little different for the ones used to the normal sandwich ELISA format.
The OD is inversely correlated to the amount of nitrotyrosine-proteins in your sample. I measured such proteins in whole tissue lysates and I had to fiddle around a bit to get an optimal sample dilution.
The ELISA is fairly straightforward. You can show the results as change of the OD over time (just set the reader to measure every few seconds) or do an end-point reading after stopping the reaction with HCl.

Abcam user community

Verified customer

Submitted Apr 08 2015

The antibody used in the Nitrotyrosine ELISA kit (ab113848), and thus the kit itself, is reactive with multiple species including rat. The antibody was prepared using nitrated KLH as immunogen and screened using nitrated BSA. It recognizes multiple nit...

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Thank you for contacting us.

The competitive ELISA is considered to be more sensitive than the sandwich ELISA. Both are compatible with your samples

Please do not hesitate to contact us if you need any more advice or information. ...

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Thank you for your message. I am sorry that this product did not work for you this time. I will look forward to contact from xxxxx. Thank you for your continued cooperation. Please do not hesitate to contact us if you have any future questions or...

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Thank you for these additional details.

Your experimental set up is good and you have done a nice job of following the steps and setting the experiment up correctly. Wecan’t say why these samples at very high concentration areincreasin...

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Thank you for contacting us.

We haven’t measured nitration in serum. Our lab wants to make sure that when using this kitthat the blocking buffer, diluents, pH and ionic strength for all samples and standards was...

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Thank you for providing this information.

I have reviewed this with my colleagues in the lab. It appears to us as if the very high protein concentrations are affecting the assay. I can’t understand why it would be a negative curve thou...

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Thank you for contacting us.

I have been in touch with the lab regarding your issue.Unfortunately we are at a loss to explain why more protein would cause a greater signal. There is, as we spoke of on the phone, the possibility that t...

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1-10 of 22 Abreviews or Q&A


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