ELISA/assay

Nitrotyrosine ELISA Kit

SKT-126

96 well

Nitrotyrosine ELISA Kit

Colorimetric detection of Nitrotyrosine

96 well

Cancer Oxidative Stress Cell Signaling Post-translational Modifications Nitration Neuroscience Neurodegeneration Alzheimer's Disease Parkinson's Disease

Nitro tyrosine ELISA Kit, 3-Nitrotyrosine ELISA Kit, 3-Nitro-L-tyrosine ELISA Kit, 3 Nitrotyrosine ELISA Kit, 3-NT ELISA Kit, 3NT ELISA Kit

ELISA Kits

Colorimetric Assay

Competitive ELISA (Enzyme-linked Immunosorbent Assay)

Competitive

ELISA kit used to quantitate Nitrotyrosine in samples.

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.

Intra-Assay Precision: Three samples of known concentration were assayed thirty times on one plate. The intra-assay coefficient of variation of the Nitrotyrosine ELISA has been determined to be <10%. Inter-Assay Precision: Three samples of known concentration were assayed thirty times in three individual assays. The inter-assay coefficient of variation of the Nitrotyrosine ELISA has been determined to be <15%.

Species Independent

Plasma Serum Cell Lysates Urine

38 samples in duplicate

1 hour

50 nM

62.5 - 8000 nM

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.

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