Nitrotyrosine ELISA Kit

Nitrotyrosine is a stable biomarker formed by the nitration of tyrosine residues in proteins, typically through the action of reactive nitrogen species such as peroxynitrite. It serves as a reliable indicator of nitric oxide (NO) -dependent oxidative damage and chronic inflammation. Unlike transient NO metabolites, nitrotyrosine accumulates in tissues and circulates in the bloodstream, offering a longer-lasting signal of nitrosative stress. In neuroscience, elevated nitrotyrosine levels have been detected in neuroinflammatory and neurodegenerative conditions, including Alzheimer’s disease, Parkinson’s disease, and multiple sclerosis. Its presence reflects oxidative damage to neuronal proteins, disruption of cellular signaling, and mitochondrial dysfunction—hallmarks of progressive neurodegeneration. Because protein-bound nitrotyrosine is more stable than free NO metabolites, it provides a more accurate measure of sustained oxidative injury in the brain. Its detection in cerebrospinal fluid and brain tissue supports its use as a diagnostic and prognostic biomarker in neurological research.

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.

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

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%.