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The multispecies 3-Nitrotyrosine (3-NT) ELISA quantitates 3-NT in serum, plasma, and other biological fluids
Principle of the method:
The Competitive multispecies ELISA research-use-only kit is designed to quantitatively measure 3-Nitrotyrosine (3-NT) independent of species. A 3-NT standard is provided to generate a standard curve for the assay and all samples are read off a user-generated standard curve. Standards or diluted samples are pipetted into a coated microtiter plate and Biotinylated Detection Ab specific to 3-NT is added. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured on a microplate reader. The concentration of 3-NT in the samples is then determined by comparing the OD of the samples to the standard curve.
Rigorous validation:
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxidesignaling. 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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