Description
CyDye DIGE fluors are available as minimal and saturation labeling dyes. The minimal dyes are intended for general 2-D application use where sufficient amounts of sample are available. The saturation dyes from the Scarce Sample Labeling Kit are designed to be used for applications where only small amounts of sample are available, for example in Laser Capture Microdissection. In total, three minimal dyes and two saturation dyes are available. CyDye DIGE fluors are exceptional dyes for multicolor analysis, offering bright and intense colors with narrow excitation and emission bands. The fluors are spectrally distinct, making them highly suitable for multicolor detection. CyDye DIGE fluors utilize these benefits but are also size- and charge-matched specifically for 2-D DIGE using Ettan DIGE system. CyDye DIGE Fluor Minimal Dyes 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.
