Nitrotyrosin (EDTA-Plasma) ELISA Assay Kit


The Eagle Biosciences Nitrotyrosin (EDTA-Plasma) ELISA Assay Kit is an enzyme immunoassay intended for the quantitative determination of protein-bound nitrotyrosine in human EDTA-plasma, serum and dried blood spots. For research use only. Not for use in diagnostic procedures.

Nitrotyrosin (EDTA-Plasma) ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Standard Range: 4.9-400 nM
Incubation Time: 1h, 1h, 10-20min
Sample Type: EDTA Plasma, Serum, Dried Blood Spots
Sample Size: 15 µl (EDTA Plasma, Serum) | 50 µl (dried blood)

Product manufactured in Germany by Immundiagnostik

Additional Information

Assay Background

Nitrotyrosine is the nitrated form of the amino acid tyrosine. The accumulation of protein bound nitrotyrosine is associated with cardiovascular diseases that are based on inflammatory processes (e.g., atherosclerosis, myocardial infarction, diabetic vasculopathy, hypertension, or coronary heart diseases). A growing number of studies have also associated the accumulation of nitrotyrosine with neurological diseases (Alzheimer´s disease, Parkinson´s disease, multiple sclerosis, stroke). With treatment of some of the associated diseases the levels of nitrated tyrosines have been shown to decrease, so nitrotyrosine has been stated to be a marker of nitrosative stress. During inflammatory processes, large amounts of nitric oxide (•NO) are locally released from L-arginine. This reaction is catalysed by the enzyme NO-synthase (NOS). Other causes for the increased •NO production are exposure to chemicals or heavy metals, drugs, nicotine, or physical and psychological stress, as well as extraordinary physical strain with increased oxygen consumption. In high concen-trations, •NO that is not trapped by mitochondrial superoxide dismutase (MnSOD) reacts with superoxide (•OO–) to form peroxynitrite. (ONOO–). Peroxynitrite is implica-ted as a key oxidant species in several pathologies and is known to be cytotoxic (nitrosative stress). Peroxinitrite is highly reactive and shows a high affinity to aromatic amino acids, e.g., to the phenolic ring of tyrosine. The nitration of tyrosine in general is a natural process within the post-translational protein modification. Nitrotyrosine is a stable product and might be seen as a correlate of peroxynitrite production, and its accumulation in cells and tissues is a marker of oxidative stress and nitrosative stress, respectively.

Assay Principle

This ELISA is designed for the quantitative determination of nitrotyrosine. The assay utilises the “sandwich” technique. Standards, controls and prepared samples which are assayed for nitrotyrosine are added into the wells of a micro plate coated with polyclonal goat anti- nitrotyrosine antibody. During the first incubation step, nitrated proteins are bound by the immo-bilised primary antibody. Then a peroxidase-conjugated polyclonal goat anti-human serum proteins antibody is added into each microtiter well and a “sandwich” of primary antibody – nitrated protein – peroxidase-conjugate is formed. Tetramethylbenzidine is used as peroxidase substrate. Finally, an acidic stop solution is added to terminate the reaction. The colour changes from blue to yellow. The intensity of the yellow colour is directly proportional to the concentration of nitrotyrosine. A dose response curve of the absorbance unit (optical density, OD at 450 nm) vs. standard concentration is generated, using the values obtained from the standards.


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