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2D DIGE vs. 2D Gel

 

 

 

Why run 2D DIGE instead of 2D gels?

In 2D-DIGE (two-dimensional difference in gel electrophoresis), we use fluorescent dyes to label proteins, can run up to 3 samples in one gel. There are many advantages to 2D-DIGE over traditional 2D gels:



2D DIGE

Standard 2D


Labeling method


Minimal dye


Staining


Spot sensitivity


0.2 ng/spot


50 ng/spot (Coomassie)
1 ng/spot (Silver)
Sypro Ruby: 1 ng/spot


Number of samples per gel


3 per gel


1 per gel


Number of in-gel comparisons


3 comparisons per 1 gel


1 comparison per 2 gels


Spot resolution


Higher


Lower


Reproducibility


Higher


Lower

 

1. High sensitivity. The CyDye fluorescent dyes have a sensitivity of 0.2 ng/spot, as compared to the sensitivity of Coomassie at 100 ng/spot. This allows us to run smaller amounts of protein on the 2D DIGE gels and results in much better spot resolution than traditional 2D gels stained by Coomassie. Scanned gels are publication quality.

2. High accuracy. The extremely high spot resolution enables accurate software-aided spot quantitation and protein expression comparison between samples. Differences in protein expression as small as 10% can be detected, and protein isoforms and post-translational modifications are easily visualized.

3. Fewer number of gels. Since the protein expression patterns from 3 different samples are compared in the same gel, fewer gels are required per project. In addition, there is no need to run technical replicates as in standard 2D.

4. Fast turn-around. Software aided in-gel analysis enables fast turn-around time, customers receive a data report within 1 week after the samples are received.

5. Cost effective. The advantages of 2D DIGE allow us to competitively price our services. The price of the 2D DIGE gel includes sample preparation (both standard and customized), protein concentration determination, CyDye labeling, isoelectric focusing, 2D SDS-PAGE, gel scanning, image preparation, quantitative in-gel analysis of up to 3 pair-wise comparisons, data report and up to 2 hours of consultation with the project leader.