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Catalog # HIT501

Nitrotyrosine, Competitive ELISA kit

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Nitrotyrosine (NO) has been identified as a marker of inflammation and NO production. Nitrotyrosine is formed in presence of the active metabolite NO. Nitrosylation of the amino acid tyrosine occurs both for free tyrosine and protein bound tyrosine. 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 nitrogen oxide species (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 serum and plasma nitrotyrosine levels are low or sometimes even undetectable.

The competitive Nitrotyrosine ELISA kit is to be used for in vitro quantitative determination of nitrotyrosine in serum, plasma and faeces samples.

The competitive Nitrotyrosine ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the inhibition principle with a working time of 3½ hours. The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are pre-incubated with biotinylated tracer antibody in U-shape microtiter plate. Pre-incubated samples and standards with the biotinylated tracer antibodies are incubated on Nitrotyrosine-HSA (NO-HSA) coated strips. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the known standards of the NO-HSA standards (log). The nitrotyrosine concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Product type
1 x 96 det.
Standard range
46.9 to 3000 nM
Detection level
46.9 nM
Working volume
100 µl/well
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Autoimmunity, Cardiology and metabolism
Safety Data Sheet Assay Manual
376 kb

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