Homemade Colloidal Coomassie Blue Staining

 Working Solution (1L):

In In a large beaker add the ingredient in the order listed.  Make sure the Ammonium Sulfate is dissolved before adding the Coomassie G-250 solution to a final of 0.1%. Bring final volume up to 1L with H20. Store at room temperature with a tightly sealed lid.

 Procedure:

 1.  After SDS PAGE rinse Gel twice with 100 ml Distilled H20 (3 minutes each wash).

2.  Add Coomassie Blue working solution to Gel and stain for at least 2 hours.

3.  Bands should be visible without de-staining but you can de-stain with H20.

 

This Protocol was modified from references:

 Kang et al. (2002).  Highly Sensitive and Fast Protein Detection with Coomassie Brilliant Blue in Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis Bull. Korean Chem. Soc. 2002, vol. 23, No. 11 1511-1512.

 Neuhoff et al. (1988). Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250.Electrophoresis. 1988 Jun;9(6):255-62.

 

 

M    Induced   Uninduced     Purified