Related Experiment Video
Updated: Jul 7, 2026
14:43
Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes
Published on: November 26, 2008
Difference gel electrophoresis (DIGE) using CyDye DIGE fluor minimal dyes
Bulbul Chakravarti1, Sean R Gallagher, Deb N Chakravarti
1Keck Graduate Institute of Applied Life Sciences, Claremont, California, USA.
Current Protocols in Molecular Biology
|February 12, 2008
Summary
Difference gel electrophoresis (DIGE) offers sensitive protein quantification. This technique uses CyDye labeling for co-detection and co-analysis, enhancing proteomic studies.
Area of Science:
- Proteomics
- Biochemistry
- Molecular Biology
Background:
- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is a standard method for protein separation and quantification.
- Traditional staining methods in SDS-PAGE can lack sensitivity and dynamic range.
- Difference gel electrophoresis (DIGE) emerged as an advanced technique to overcome these limitations.
Purpose of the Study:
- To introduce and explain the principles of Difference Gel Electrophoresis (DIGE) as an advanced proteomic technique.
- To highlight the advantages of DIGE, including its sensitivity and wide dynamic range.
- To discuss the applications of DIGE in both one-dimensional (1-D) and two-dimensional (2-D) SDS-PAGE for quantitative proteomic analysis.
Main Methods:
- Utilizes fluorescent CyDye minimal dyes for differential labeling of protein samples.
- Involves combining differentially labeled samples before electrophoresis.
- Employs co-detection and co-analysis on the same gel for comparative quantitative measurements.
Main Results:
- DIGE provides highly sensitive protein detection with a broad dynamic range.
- The technique allows for precise quantitative comparisons between different protein samples.
- DIGE is compatible with subsequent protein identification using mass spectrometry.
Conclusions:
- DIGE represents a significant advancement over traditional SDS-PAGE for quantitative proteomics.
- The use of CyDye labeling and co-analysis enhances the accuracy and sensitivity of protein expression studies.
- Both 1-D and 2-D DIGE are valuable tools for comprehensive proteomic investigations.
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...
02:35DNA Agarose Gel Electrophoresis
Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
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,...
01:20Electrophoresis: 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...