RPK0272
            
CYDYE DIGE CY2 MINIMAL 25 NMOL
        
        

            
RPK0273
            
CYDYE DIGE CY3 MINIMAL 25 NMOL
        
        

            
RPK0275
            
CYDYE DIGE CY5 MINIMAL 25 NMOL
        
        

            
RPK0283
            
CY3 CYSTEINE SAT. DYE 2D DIGE
        
        

            
RPK0285
            
CY5 CYSTEINE SAT. DYE 2D DIGE
        
        

            
25-8008-60
            
CYDYE DIGE CY2 MINIMAL 10 NMOL
        
        

            
25-8008-61
            
CYDYE DIGE CY3 MINIMAL 10 NMOL
        
        

            
25-8008-62
            
CYDYE DIGE CY5 MINIMAL 10 NMOL
        
        

            
25-8009-83
            
CYDYE DIGE SCARCE SAMPLE
        
        

            
25-8009-84
            
CYDYE DIGE SCARCE SAMPLE+PREP
        
        

            
25-8010-65
            
5NMOL CYDYE DIGE FLUOR MIN KIT
        
        

            
25-8010-82
            
CYDYE DIGE CY2 MINIMAL 5 NMOL
        
        

            
25-8010-83
            
CYDYE DIGE CY3 MINIMAL 5 NMOL
        
        

            
25-8010-85
            
CYDYE DIGE CY5 MINIMAL 5 NMOL
        
        

            
28-9345-30
            
CyDye DIGE Fluor, minimal labe
        
        

            
28-9366-83
            
CyDye DIGE PREP SCARCE SAMPLE
        
    


CyDye™ DIGE Fluors

    
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.

CyDye™ DIGE Fluors
Technical Information


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 ideal 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 subject 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.
 
The NHS ester reactive group of CyDye DIGE Fluor minimal dyes covalently attaches to the epsilon amino group of lysine of proteins via an amide linkage. The ratio of dye to protein has been specifically designed to ensure the dyes are limiting in the reaction. As a result, approximately 3% of the available proteins are labeled and then only on a single lysine per protein (i.e., one dye per protein, or minimal labeling).
 
The amino acid lysine in proteins carries an intrinsic single positive charge at neutral or acidic pH. CyDye DIGE Fluor minimal dyes also carry a single positive charge which, when coupled to the lysine, replaces the single positive charge of the lysine with its own, ensuring that the pI of the protein does not significantly alter compared with the same unlabeled protein.
 
CyDye DIGE Fluor Labeling Kits for Scarce Samples

Two labeling kits for scarce samples are available: one contains the Cy™3 and Cy5 saturation dyes; the other contains those and an additional vial of Cy3 dye to label a preparative gel. Each kit contains sufficient dye for at least 12 labeling reactions and allows labeling of as little as 5 μg protein per labeling reaction. The CyDye DIGE Fluor Labeling Kit for Scarce Samples and Preparative Gel Labeling contains an additional vial of Cy3 dye for labeling up to 500 μg of protein. The saturation dyes Cy3 and Cy5 from the Labeling Kit for Scarce Samples retain the advantages described for the minimal dyes. The saturation dyes in the two kits allow the labeling of 5 μg protein per labeling reaction compared to 50 μg with the minimal dyes. The maleimide reactive group of the saturation dyes covalently bonds to the thiol group of cysteine residues of proteins via a thioether linkage.
 
To achieve maximal labeling of cysteine residues, the protocol uses a high fluor to protein labeling ratio. This type of labeling method labels all available cysteines on each protein under the conditions used, resulting in the majority of cysteine groups in a protein from a sample being labeled. For this reason, the method has been called "saturation labeling."
 
The CyDye DIGE Fluor saturation dyes covalently label cysteine residues. They have a net neutral charge and therefore do not alter the charge on a protein after labeling. They are also matched for molecular weight ensuring that the same protein from a standard and test sample labeled with the different dyes will overlay when separated on a single gel.
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