17-376 | Nitrotyrosine ELISA

17-376
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      Overview

      Key Spec Table

      Analytes Available Species Reactivity Key Applications Detection Methods
      NitrotyrosineCa,M,H,R,Po,Eq,Rb,Gp,Ht,NhPELISAChemiluminescence
      Description
      Catalogue Number 17-376
      Brand Family Upstate
      Trade Name
      • Upstate
      Description Nitrotyrosine ELISA
      Overview Oxidative stress has been implicated in the pathogenesis of neurodegenerative disorders, cancer, and aging. Oxidative stress occurs as a response to increased oxidants, decreased anti-oxidants, or failure to repair oxidative damage induced by reactive oxygen species (ROS). ROS are free radicals, reactive molecules containing oxygen, or molecules containing oxygen that generate free radicals. ROS include nitric oxide (NO), superoxide (O2-), peroxynitrite (ONOO-), and hydroxyl radical (OH-). ROS are cleared from cells by the action of superoxide dismutase (SOD), catalase and peroxidases.

      NO, produced by nitric oxide synthases (NOS), is a critical signaling molecule involved in many physiological processes including muscle contraction, vasodilation, neuronal transmission, and immune responses. Some of these effects are mediated through reaction with and activation of guanylate cyclase (GC), resulting in increased cGMP production. With a half-life of seconds, NO readily diffuses through the cytosol and cell membrane, allowing it to act inside or adjacent to the cell in which it was produced.

      ROS may interact to form agents that modify cellular proteins. Superoxide, produced by NADPH oxidase (phox), reacts with NO to form peroxynitrite. Intracellular peroxynitrite can modify proteins by interacting with and nitrating tyrosine residues to form 3-nitrotyrosine. Tyrosine nitration may increase (e.g. sGC, Src, PI3K, Akt), decrease (e.g. Mn-SOD, Ca++-ATPase), or have no discernable effect (e.g. p53, VASP, α-Synuclein) on the activity of a particular protein.

      Upstate's new Nitrotyrosine Assay Kit, Chemiluminescence Detection (Cat. # 17-376) is a competitive ELISA for the quantitation of tyrosine nitration. The kit includes all reagents needed for measurement of tyrosine nitration, including white high binding 96-well plates, nitrated BSA standard, a Nitrotyrosine antibody, LumiGLO® chemiluminescent detection substrate, and wash buffers.

      The assay has a wide dynamic range and high precision, making this assay a valuable new tool for the study of tyrosine nitration.
      References
      Product Information
      Components LumiGLO® Chemiluminescent Substrate. Reagent B. Catalog # 20-212-d. One vial containing 10 mL of substrate. White high binding plates. Two Costar 3922 white high-binding 96-well plates. TBS, 20X. Catalog # 20-190C. One vial containing 100 mL of 1M Tris, 3M NaCl, pH 7.4. 20% Tween®-20 (v/v). Catalog # 20-246. One vial containing 3 mL of 20% Tween®-20 in sterile, distilled water. Blocking Buffer 2, 10X. Catalog # 20-316. One vial containing 7.5 mL of a proprietary buffer formulation, 10X stock containing 0.05% Kathon®. Anti-Nitrotyrosine, 200X. Catalog # 07-752. One vial containing immuno-affinity purified rabbit IgG in 25mM Tris, pH 7.4, 137mM sodium chloride, 2.7mM potassium chloride, 0.05% Kathon® and 5mg/ml BSA, 200X stock. Nitrated BSA. Catalog # 13-127. One vial containing 1 mg in 200 mL of nitrated BSA in 25mM Tris, pH 7.4, 137mM sodium chloride, 2.7mM potassium chloride. The 3-nitrotyrosine content was determined spectrophotometrically (ξ 438 =4300 M-1 cm-1) to be 5.6 mol nitrotyrosine per mol BSA. Goat Anti-Rabbit IgG, HRP-conjugate. Catalog # 12-348-MN. One vial containing 25 μg goat anti-rabbit IgG conjugated to horseradish peroxidase, in 25 μL 0.02M potassium phosphate, 0.15M NaCl, pH 7.2, 10 mg/mL BSA, 0.01% (w/v) gentamicin sulfate before the addition of glycerol to 50%. LumiGLO® Chemiluminescent Substrate. Reagent A. Catalog # 20-212-c. One vial containing 10 mL of substrate.
      Detection method Chemiluminescence
      Applications
      Application This Nitrotyrosine ELISA is used to measure & quantify Nitrotyrosine levels in Neuroscience & Toxicity research.
      Key Applications
      • ELISA
      Biological Information
      Species Reactivity Canine Mouse Human Rat Pig Horse Rabbit Guinea Pig Hamster Non-human Primate
      Analytes Available
      • Nitrotyrosine
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Packaging Information
      Material Size 1 kit
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Languages
      English

      Certificates of Analysis

      TitleLot Number
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2113321 2113321
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2139667 2139667
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2382114 2382114
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2441919 2441919
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2445168 2445168
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2023293 2023293
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2025138 2025138
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2178408 2178408
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2207263 2207263
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2239864 2239864
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - 2281657 2281657
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1460106 DAM1460106
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1472137 DAM1472137
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1487517 DAM1487517
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1518791 DAM1518791
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1527883 DAM1527883
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1548608 DAM1548608
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1570850 DAM1570850
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1598745 DAM1598745
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1658648 DAM1658648
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1745425 DAM1745425
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1764561 DAM1764561
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1785939 DAM1785939
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - DAM1805305 DAM1805305
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - JBC1389846 JBC1389846
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - JBC1863292 JBC1863292
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - JBC1889918 JBC1889918
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - JBC1926738 JBC1926738
      Nitrotyrosine Assay Kit, Chemiluminescence Detection - JBC1946434 JBC1946434
      Nitrotyrosine Assay Kit, Chemiluminescence Detection -2510240 2510240
      Nitrotyrosine Assay Kit, Chemiluminescence Detection -2512423 2512423

      References

      Reference overviewPub Med ID
      Low-level laser light therapy improves cognitive deficits and inhibits microglial activation after controlled cortical impact in mice.
      Jugta Khuman,Jimmy Zhang,Juyeon Park,James D Carroll,Chad Donahue,Michael J Whalen
      Journal of neurotrauma 29 2012

      Show Abstract
      21851183
      Hydrogen gas reduced acute hyperglycemia-enhanced hemorrhagic transformation in a focal ischemia rat model.
      C H Chen,A Manaenko,Y Zhan,W W Liu,R P Ostrowki,J Tang,J H Zhang
      Neuroscience 169 2010

      Show Abstract Full Text Article
      20423721
      Free radical stress-mediated loss of Kcnj10 protein expression in stria vascularis contributes to deafness in Pendred syndrome mouse model.
      Ruchira Singh,Philine Wangemann
      American journal of physiology. Renal physiology 294 2008

      Show Abstract
      17959752

      Data Sheet

      Title
      Nitrotyrosine ELISA - Data Sheet