Analysis Note

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Application

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.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.

Features and Benefits

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.

General description

CyDye^™ DIGE Fluor Preparative Gel Labeling for Scarce Samples, Cy3^™ saturation dye (400 nmol).CyDye^™ DIGE Fluor minimal dyes and saturation dyes are size- and charge-matched fluorescent dyes specifically designed for detecting protein abundance differences in 2-D Fluorescence Difference Gel Electrophoresis using Ettan^™ DIGE system.

Legal Information

Ettan is a trademark of Cytiva

Cy3 is a trademark of Cytiva

Immobiline is a registered trademark of Cytiva

CyDye is a trademark of Cytiva