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Validation of Different Staining Methods of the 2D Gels

 

 

 

In order to validate the sensitivity of CyDye labeling, we performed a side-by-side comparison of  the 2D gel images from CyDye-labeling, Silver staining and Blue Silver Staining (an improved Coomassie staining). The data clearly indicates:

  • CyDye labeling is more sensitive than Silver staining
  • Blue Silver staining is more accurate than Silver staining
  • CyDye labeling, Silver staining, and Blue Silver staining method generate the same protein pattern

 

CyDye Labeling Detects More Spots Than Silver Staining Using The Same Gel

Gel-1: Silver staining*

Coomassie Staining of 2D Gels

Gel-1 : Cy2-labeling*

Cy3 labeling of 2D gels

* Images are from the same gel, spots with red circles are clearly detectable with CyDye labeling but not with Silver Staining.


Blue Silver Staining Detects More Spots Than Silver Staining (E.coli: pH3-10NL)

Gel-1: Silver Staining

Deep Purple  Staining of 2D Gels
Gel-2: Blue Silver Staining

Cy5 labeling of 2D gels


* Spots with red circles are clearly detectable with Blue Silver Staining but not with Silver Staining.

CyDye Labeling and Blue Silver Staining Are Comparable Using The Same Gel

Gel-2: Blue Silver staining*

Luminteine Staining of 2D Gels

Gel-2 : Cy2-labeling*

Sypro Ruby  Staining of 2D Gels

* Images are from the same gel, spots with white circles are clearly detectable with both Blue Silver Staining and CyDye labeling.


In addition, we also compared CyDye labeling with Other Staining Methods such as Deep Purple, Sypro Ruby, Luminteine and regular Commassie staining, the data clearly indicates:

  • CyDye labeling is slightly more sensitive than Deep Purple, Sypro Ruby, Luminteine, Silver Staining
  • CyDye labeling is much more sensitive than regular Coomassie Staining
  • CyDye labeling, Deep Purple, Sypro Ruby, Luminteine Staining and Commmassie Staining generate the similar protein pattern