2D DIGE vs 2D Gel

Why run 2D DIGE instead of 2D gel electrophoresis?

In 2D gel, proteins are separated by PI (isoelectric point) in IEF (1st dimension) and molecular weight in SDS PAGE (2nd dimension).

2D DIGE is an improved version of standard 2D gel electrophoresis. In addition to all the features of 2D gel, 2D DIGE labels different samples with up to 3 fluorescent tags. This allows up to 3 samples to be run on the same gel—saving cost, minimizing gel-to-gel variation, and having higher sensitivity/accuracy.

2D DIGE vs 2D Gel experimental procedure
2D DIGEStandard 2D Gel
Staining MethodFluorescent DyeStaining
Spot Sensitivity0.2 ng/spotCoomassie blue: 50 ng/spot
Silver staining: 1 ng/spot
Sypro Ruby: 1 ng/spot
Number of samples per 2D gel3 per gel1 per gel
Number of in-gel comparisons3 comparisons per gel1 comparison per 2 gels
Spot resolutionHigherLower

Advantages of 2D DIGE over 2D Gel electrophoresis:

  • Higher sensitivity: 2D DIGE uses Fluorescent labeling with a sensitivity of 0.2 ng/spot, versus the sensitivity of standard 2D gel electrophoresis with Coomassie blue staining at 100 ng/spot or silver staining at 1 ng/spot
  • Higher accuracy: The extremely high spot resolution enables accurate spot quantitation. Differences in protein expression as small as 10% can be detected
  • Higher reproducibility: Nearly identical data was obtained on the same sample labeled with different CyDye on the same gel or cross different gels, thus eliminating the need of running technical replicates
  • Broader spectrum: Our large 2D DIGE gel format resolve ~5000 protein spots, allowing a wide dynamic range detection/quantitation of low abundant proteins, large proteins and small peptides
  • Fewer number of gels: Each 2D DIGE gel can accommodate up to 3 different samples, versus 1 sample on each 2D Gel. For the same number of samples, only 1/3 number of 2D DIGE gels is needed.
  • Lower cost: Our price is based on each gel instead of each sample. Lower number of 2D DIGE gels and no need of technical replicates translates into higher quality data at a lower cost than standard 2D Gel service. Price covers from sample preparation to publication-ready data.
  • Fast turn-around: Results In 5-7 Days.