Related Experiment Video
Updated: Jul 20, 2026
12:07
Profiling Thiol Redox Proteome Using Isotope Tagging Mass Spectrometry
Published on: March 24, 2012
Detection of redox-based modification in two-dimensional electrophoresis proteomic separations
1Department of Biochemistry, University College Cork, Lee Maltings, Prospect Row, Mardyke, Cork, Ireland. d.Sheehan@ucc.ie
Biochemical and Biophysical Research Communications
|September 8, 2006
Summary
Oxidative stress damages cells by altering proteins. Two-dimensional electrophoresis (2D SDS-PAGE) is a key proteomic method for identifying these redox-based protein modifications and understanding their effects.
Area of Science:
- Biochemistry
- Proteomics
- Cell Biology
Background:
- Oxidative stress occurs when reactive oxygen species (ROS) overwhelm cellular antioxidant defenses.
- ROS can lead to detrimental modifications of lipids, DNA, and proteins.
- Understanding protein modifications is crucial for dissecting oxidative stress impacts.
Purpose of the Study:
- To review methodologies for identifying redox-based protein modifications.
- To highlight the utility of 2D SDS-PAGE in oxidative stress research.
- To explore the implications of identifying specific protein targets.
Main Methods:
- Utilizing two-dimensional electrophoresis (2D SDS-PAGE) for protein separation.
- Analyzing redox-based modifications including carbonylation, glutathionylation, and disulphide bridge alterations.
- Employing proteomic analysis for protein identification.
Main Results:
- 2D SDS-PAGE enables rapid, quantitative analysis of protein modifications.
- Diverse reversible and irreversible alterations to amino acid side chains are observed.
- Identification of modified proteins provides insights into cellular responses.
Conclusions:
- 2D SDS-PAGE is a robust platform for studying oxidative stress-induced protein modifications.
- Identifying specific protein targets deepens the understanding of oxidative stress mechanisms.
- These findings have potentially far-reaching implications for cellular biology and disease.
More Related Videos
10:51Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting
Published on: April 10, 2014
12:34Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome
Published on: April 2, 2018
Related Concept Videos
01:22Two-dimensional Gel Electrophoresis
Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
01:20Electrophoresis: Overview
Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...
There...
01:27SDS-PAGE
Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
01:30Capillary Electrophoresis: Applications
Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
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.