What is DiGE?
Difference Gel Electrophoresis (DiGE) is a modification of 2-D PAGE. Two (see figure below – from 2-D Electrophoresis. Principles and Methods; GE Healthcare) or three separate protein samples are labeled with different fluorescent dyes prior to separation, enabling accurate analysis of differences in protein abundance between samples.Schematic DiGE analysis  |
We use CyDye TM DIGE Fluor dyes (Cy2, Cy3, and Cy5). These dyes show the following characteristics:
| Size- and charge matched | The same labeled protein from different samples will migrate to the same position, regardless of the dye used |
| pH insensitive | No change in signal over the wide pH range used during first-dimension separation (IEF) and equivalent migration in SDS gels |
| Spectrally resolvable | The distinct signal from each fluor contributes to the accuracy |
| Highly sensitive and bright | As little as 125 pg of protein can be detected |
| Photostable | There is minimal loss of signal during labeling, separation, and scanning |