2D DIGE vs Silver Staining

How does 2D DIGE compare to Silver Staining?

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 sensitive than Silver staining

CyDye labeling detects more spots than silver staining

Gel-1: Silver staining

2D Gel with silver staining: spots in red circles are detected by CyDye labeling but not silver staining

Gel-1: Cy2-labeling

2D Gel with CyDye labeling: spots in red circles are detected by CyDye labeling but not silver staining

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

2D Gel with silver staining: spots in red circles are detected by CyDye labeling but not silver staining

Gel-2: Blue Silver staining

2D Gel with blue silver staining: spots in red circles are detected by blue silver but not silver staining

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

CyDye labeling and blue silver staining are comparable

Gel-2: Blue Silver staining

2D Gel with blue silver staining: spots in white circles are detected by blue silver staining and CyDye labeling

Gel-2 : Cy2-labeling

2D Gel with blue CyDye labeling: spots in white circles are detected by CyDye labeling and blue silver staining

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