Related Experiment Video
Updated: May 17, 2026
07:32
Detection of 3-Nitrotyrosine in Atmospheric Environments via a High-performance Liquid Chromatography-electrochemical Detector System
Published on: January 30, 2019
Detection of nitrosated proteins.
1Boston University School of Medicine, Boston, Massachusetts, USA.
Current Protocols in Toxicology
|October 10, 2012
Summary
This study presents three methods for detecting nitrosated proteins. The Saville assay is common, while diaminonaphthalene and chemiluminescence assays offer high sensitivity for protein nitrosation detection.
Area of Science:
- Biochemistry
- Analytical Chemistry
Background:
- Nitrosated proteins are important biomarkers in various physiological and pathological processes.
- Accurate detection methods are crucial for understanding protein nitrosation.
Purpose of the Study:
- To outline and compare methods for detecting nitrosated proteins.
- To guide researchers in selecting appropriate detection techniques based on experimental needs.
Main Methods:
- The Saville assay, a spectrophotometric method, is described as a common and robust technique.
- Fluorescence-based methods, including the diaminonaphthalene assay, are presented for high-sensitivity applications.
- Chemiluminescence assays are also highlighted for sensitive detection of nitrosated proteins.
- Mass spectrometry is mentioned for precise structural characterization of nitrosated proteins.
Main Results:
- The Saville assay offers advantages in reduced interference and use of simple reagents.
- Diaminonaphthalene and chemiluminescence assays provide enhanced sensitivity.
- Mass spectrometry enables definitive structural identification.
Conclusions:
- Multiple methods exist for nitrosated protein detection, each with specific advantages.
- Method selection depends on the required sensitivity and structural information.
- These methods are essential tools in biochemical research and diagnostics.
Related Concept Videos
01:202° Amines to N-Nitrosamines: Reaction with NaNO2
Secondary amines react with nitrous acid to form N-nitrosamines, as depicted in Figure 1. Nitrous acid, a weak and unstable acid, is formed in situ from an aqueous solution of sodium nitrite and strong acids, such as hydrochloric acid or sulfuric acid, in cold conditions. In the presence of an acid, the nitrous acid gets protonated. The subsequent loss of water results in the formation of the electrophile known as nitrosonium ion.
01:15Western Blotting
Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.