DIGE (Differential Gel Electrophoresis) is designed to provide a quantitative component to proteomics experiments utilizing two-dimensional (2D) gel electrophoresis. DIGE can provide detection of changes in protein abundance (sometimes subtle) with statistical confidence while controlling for gel-to-gel variation and other variations of non-biological origin. Samples (controls and treated) are differentially labeled with spectrally resolvable fluorescent dyes (Cy2, Cy3 and Cy5) and co-resolved on a single 2D gel for direct quantitation. Using internal standards and experimental repetition, single and multi-variant analyses can be inter-compared with a relatively small number of coordinated DIGE gels.
For more detailed information regarding DIGE projects please see our DIGE Handout and for more technical information regarding the DIGE technology please visit GE Healthcare.
A flowchart of the process:
- A planning meeting with the Investigator
- Sample Preparation (Extraction and clean-up)
- Labeling (Cy2, Cy3 & Cy5)
- Analytical 2D gel electrophoresis
- Laser Imaging (Typhoon Imager)
- Computerized Image Analysis (DeCyder software)
Please contact the PCL for additional information about DIGE. Careful planning must be given to acheive maximal success.