Unit

UNIT 17.15 High-Throughput, Multiplexed Analysis of 3-Nitrotyrosine in Individual Proteins

  1. Hongjun Jin,
  2. Richard C. Zangar

Published Online: 1 FEB 2012

DOI: 10.1002/0471140856.tx1715s51

Current Protocols in Toxicology

Current Protocols in Toxicology

How to Cite

Jin, H. and Zangar, R. C. 2012. High-Throughput, Multiplexed Analysis of 3-Nitrotyrosine in Individual Proteins. Current Protocols in Toxicology. 51:17.15:17.15.1–17.15.16.

Author Information

  1. Fundamental & Computational Sciences, Pacific Northwest National Laboratory, Richland, Washington

Publication History

  1. Published Online: 1 FEB 2012
  2. Published Print: FEB 2012

Abstract

Reactive nitrogen species (RNS) and reactive oxygen species (ROS) are derived as a result of inflammation and oxidative stress and can result in protein modifications. As such, these protein modifications are used as biomarkers for inflammation and oxidative stress. In addition, modifications in single-tissue-associated proteins released into blood can provide insight into the tissue localization of the inflammation or oxidative stress. We have developed an enzyme-linked immunosorbent assay antibody microarray platform to analyze the levels of 3-nitrotyrosine in specific proteins in a variety of biological samples, including human plasma and sputum. Selective-capture antibodies are used to immunoprecipitate individual proteins from samples onto isolated spots on the microarray chips. Then, a monoclonal antibody for 3-nitrotyrosine is used to detect the amount of 3-nitrotyrosine on each spot. Our studies suggest that this approach can be used to detect trace amounts of 3-nitrotyrosine in human plasma and sputum. In this paper, we describe our antibody microarray protocol for detecting 3-nitrotyrosine in specific proteins. Curr. Protoc. Toxicol. 51:17.15.1-17.15.16. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • 3-nitrotyrosine;
  • ELISA microarray;
  • multiplex;
  • biomarker