Nitrotyrosine ELISA Assay Kit


The Eagle Biosciences Nitrotyrosine ELISA Assay Kit is an enzyme immunoassay intended for the quantitative determination of protein-bound nitrotyrosine in EDTA plasma, serum and stool. For research use only.

SKU: KR7824 Categories: , ,

Nitrotyrosine ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Standard Range: 55.6-1500 nM
Incubation Time: 2.5h, 1h, 10-20min
Sample Type: Stool, Serum, EDTA-Plasma
Sample Size: 100 mg (Stool); 50 µl (Ser.; Pl.)

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 catalyzed 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 concentrations, •NO that is not trapped by mitochondrial superoxide dis-mutase (MnSOD) reacts with superoxide (•OO–) to form peroxynitrite (ONOO–). Per-oxynitrite is implicated 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 pro-cess 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. It utilises the sandwich technique with two polyclonal antibodies against nitrated proteins.
Standards, controls and diluted samples which are assayed for nitrotyrosine are ad-ded into the wells of a microtiter plate coated with polyclonal anti-nitrotyrosine an-tibody. During the first incubation step, nitrated proteins are bound by the immobi-lised primary antibody. Then a peroxidase-conjugated polyclonal anti-nitrotyrosine 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. concentration is generated, using the values obtained from the standards. Nitrotyrosine, present in the patient samples, is determined directly from this curve.


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