Nitrotyrosine ELISA Assay Kit


The Nitrotyrosine ELISA Assay Kit is a competitive assay that can be used for the quantification of Free Nitrotyrosine in plasma, serum, cell lysates, urine, and other sample matrices. The ELISA utilizes an Nitrotyrosine-coated plate and an HRP-conjugated antibody for detection which allows for an assay range of 62.5 to 8000nM Free Nitrotyrosine, with a sensitivity of 50nM. The other highlights of this kit are a quick incubation time of 60 minutes, stable reagents, and an easy to use protocol.

Nitrotyrosine ELISA Assay Kit

The Nitrotyrosine ELISA Assay Kit is For Research Use Only

Sensitivity: 50 nM
Dynamic Range: 62.5 – 8000 nM
Incubation Time: 1 hour
Sample Type: Cell lysates, Plasma, Serum, Urine
Sample Size: 50 μL

Product manufactured in Canada by StressMarq.

Assay Background

Nitrotyrosine has been identified as a marker of inflammation and NO production. Nitrotyrosine is formed in presence of the active metabolite NO. Various pathways including the formation of peroxinitrite lead to nitrotyrosine production. Since nitrotyrosine is a stable end product of peroxynitrite oxidation, assessment of its plasma concentration may be useful as a marker of NO-dependent damage in vivo. Since NOX is only an indicator for enhanced NO production, protein associated nitrotyrosine might be a more suitable marker for damage induced by reactive nitrogen intermediates derived from NO. Furthermore, most proteins have a longer half life in the circulation than NOX levels. The presence of nitrotyrosine has been detected in various inflammatory processes including atherosclerotic plaques, celiac disease, rheumatoid arthritis, chronic renal failure and septic shock. In normal plasma low, undetectable, levels of nitrotyrosine are present. Nitrosylation of the amino acid tyrosine occurs both for free tyrosine and for protein bound tyrosine.

Storage and Stability
This kit will perform as specified if the components are stored as directed and used before the expiration date indicated on the outside of the box. All reagents are stable as supplied at 4°C. For optimum storage, the Standard should be aliquotted into smaller portions and then stored appropriately. Avoid repeated freeze/thaw cycles (10 µL of Standard can prepare a triplicate standard curve.

Related Products

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Additional Information

Assay Procedure

  1. Prepare standard and samples in the Sample and Standard Diluent.
  2. Add 50 µL of prepared standards and samples in triplicate to appropriate wells.
  3. Add 50 µL of the diluted antibody preparation to the appropriate wells.
  4. Cover plate with Plate Cover and incubate at room temperature (20-25°C) for 1 hour.
  5. Wash plate 4 times with 1X Wash Buffer.
  6. Add 100 µL of TMB Substrate to each well.
  7. Cover plate and develop the plate in the dark at room temperature for 30 minutes.
  8. Add 100 µL of Stop Solution to each well.
  9. Measure absorbance on a plate reader at 450 nm.
  10. Plot the standard curve and calculate sample concentrations.

Typical Standard Curve

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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