3-NT(3-Nitrotyrosine) ELISA Kit (E-EL-0040)

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Universal 3-NT. During the reaction, Universal 3-NT in the sample or standard competes with a fixed amount of Universal 3-NT on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal 3-NT. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Universal 3-NT in tested samples can be calculated by comparing the OD of the samples to the standard curve.

3-nitrotyrosine modification of proteins is a wellestablished marker of protein damage by oxidative stress. 3-nitrotyrosine is a product of protein tyrosine nitration resulting fromoxidative damage to proteins by peroxynitrite. Peroxynitrite is a formedin vivo by the reaction of nitric oxide, a cellular messenger, andsuperoxide, the majority of which is generated by the mitochondrialrespiratory chain. 3-nitrotyrosine modification of proteins can result inchanges in protein structure, function and catalytic activity. Tyrosinenitration may increase (e.g. sGC, Src, PI3K, Akt), decrease (e.g. MnSOD, Ca++-ATPase), or have no discernable effect (e.g. p53, VASP, αSynuclein) on the activity of a particular protein. Tyrosine nitration hasbeen implicated in the pathogenesis of major neurological(Alzheimer's, Parkinson's, multiple sclerosis, and stroke) andcardiovascular (atherosclerosis, myocardial infarction, coronary arterydisease, hypertension) diseases that share inflammation as acontributor to pathogenesis.