3-Nitrotyrosine ELISA Kit (ab116691)

3-nitrotyrosine is a chemical formed in vivo by the reaction of tyrosine with nitrating oxidants, superoxide and nitric acid (http://www.ncbi.nlm.nih.gov/pubmed/1416975) with the chemical formula C9H10N2O5. It's structure does not change between species and is therefore species independent hence why no species have been listed on the datasheet as all species including human should be compatible with this kit. What is important is the sample type. This kit has only been tested and therefore guaranteed for use with cell culture extracts and tissue extracts.

Thank you for your enquiry.



Lung is a great sample to run with this assay!


My general recommendation is to homogenize and then sonicate the tissue. Sonicate until the sample is not sticky - DNA released from ruptured nuclei is viscous and sufficient sonication will break the DNA strands to fragments that no longer make the sample viscous. At the same time, don’t over-sonicate which can cause irreversible protein precipitation. Different samples should be normalized by BCA assay (or equivalent) for protein concentration. If there are concerns over whether or not the sample prep is appropriate, spike the kit reference protein into a sample to check for signal.



Hope this helps! Please let us know if you have further questions.

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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.

Thank you for your reply.


Our recommendation would be to collect samples using the extraction buffer provided with the kit. Without knowing the recipe of the buffer it is impossible to say whether or not it is compatible. Please let me know if this improves your results.


I wish you the best of luck with your experiments!

Thank you for your reply.

Regarding your results - can you provide the formulation of the lysis buffer you are using? As the controls and standards are working, I expect the problem is with the sample preparation. Can you also provide your data? I will have the lab investigate this further.


I look forward to your reply so that I may assist you further. Please do not hesitate to contact me if you have any additional questions.

Thank you for contacting Abcam regarding ab116691.

Unfortunately we do not sell these components separately. The kits are designed to provide sufficient amounts of reagents for 1 96 well plate, so in order to acquire additional strips, you would need to purchase another kit.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Thank you for your enquiry regarding ab116691.

3-nitrotyrosine BSA standard is from purified BSA.

I hope this information helps you, and please do not hesitate to contact us if you have further questions.

Thank you for contacting us.

Here is the response from the lab:

As far as we know we haven’t tested this kiton plasma samples. There is no reason that it is not possible, but the question is primarily one of sensitivity (are there sufficient nitrotyrosine modifications present in plasma).

If the customer wants to give it a go, suggest that they mix 1 part plasma with 4 parts extraction buffer and then prepare a dilution series from this material in incubation buffer. The kit contains 3NT BSA as a positive control so the customer will know if the assay is working even if they do not get signal from the plasma samples.

One more note on this question:We don’t know if there is anything in plasma that might interfere with this assay. Hence lack of signal with plasma samples may mean either that the nitrotyrosine in not detectable in plasma (not present or the assay is not sensitive enough) or that plasma is not suitable to apply to the assay (interferes in some way). One approach to address the later is for the customer to spike the plasma samples with the NT-BSA control. If the NT-BSA gives comparable signal +/- plasma then the assay is compatible with plasma and it comes down to a question of sensitivity.


If you wanted to try the kit on plasma samples, you would be eligible for our testing discount program. See details below.In returnyou would receive afree antibody on a future order.

For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein/kit on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.