CyDye DIGE Fluor minimal dyes are size and charge matched fluorescent dyes specifically designed for detecting protein abundance differences in 2D fluorescence difference gel electrophoresis using Ettan DIGE system. The minimal dyes are intended for general 2D application use where sufficient amounts of sample are available.
Protein samples and the internal standard are each labeled with one CyDye DIGE Fluor minimal dye. These labeled samples are then combined, run on an isoelectric focusing gel in the first dimension, and separated by SDS-PAGE in the second dimension. Electrophoresis is simplified with Ettan IPGphor 3, or Multiphor II with Immobiline DryStrip gels in the first dimension and Ettan DALTtwelve or Ettan DALTsix electrophoresis systems in the second dimension.
The ability to multiplex different CyDye DIGE Fluor minimal dye-labeled samples on the same gel means that the different samples will be subjected to exactly the same first- and second-dimension running conditions. Consequently, the same protein labeled with any of the CyDye DIGE Fluor minimal dyes and separated on the same gel will migrate to the same position on the 2-D gel and overlay. This limits experimental variation and ensures accurate within-gel matching.
- Exceptional dyes for multicolor analysis, offering bright and intense colors with narrow excitation and emission bands
- Fluors are spectrally distinct, making them highly suitable for multicolor detection
- Allow detection of up to three prelabeled protein samples and standards on the same 2-D electrophoresis gel
- Size- and charge-matched dyes enable co-migration of labeled samples within the gel
- Bright and highly sensitive dyes allow the use of the minimal labeling technique
- Minimal loss of signal during labeling, separation, and scanning
- No change in signal over wide pH range used during first-dimension (IEF) separation
- Discrete signal from each fluor with minimal cross-talk contributes to high accuracy
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