Coomassie Staining SDS-PAGE Gels

Standard Method

  1. Place gel in Staining Solution. Shake slowly for 1 hour to overnight.

  2. Replace the Staining Solution with Destaining Solution I. Shake slowly for 30 minutes.

  3. Remove Destaining Solution I and replace with Destaining Solution II. Addition of Kimwipes to one corner of the staining tray will help remove Coomassie blue from the gel without changing the destaining solution. Replace tissues when they are saturated with Coomassie blue.

  4. To minimize cracking, add 1% glycerol to the last destain before drying the gel.

Rapid Method

  1. Place gel in staining solution in a small box with a lid. (An empty pipet tip box works well).

  2. Microwave on high for 1 minute, then shake 10-20 minutes.

  3. Replace staining solution with Destain I, and add some Kimwipes or a folded-up paper towel to help absorb the stain.

  4. Microwave on high for 1 minute and shake until the bands emerge clearly from the background.

  5. Pour out Destain I, replace with distilled water. The gel will continue to destain a little bit.

  6. Dry down the gel when you get around to it.


Staining Solution
(0.025% Coomassie Brilliant blue R 250, 40% methanol,7% acetic acid)

TO MAKE 2L: 

  • 0.5 g Coomassie Brilliant blue R
  • 800 ml methanol
    Stir until dissolved. Then add:
  • 140 ml acetic acid
  • distilled H20 to 1L


Destaining Solution I
(40% methanol, 7% acetic acid)

TO MAKE 1L: 

  • 400 ml methanol
  • 70 ml acetic acid
  • distilled H20 to 1L


Destaining Solution II
(5% methanol, 7% acetic acid)

TO MAKE 10L: 

  • 500 ml methanol
  • 700 ml acetic acid
  • distilled H20 to 10L