EDC Impressum & DSE

Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Hot Coomassie Staining

April 28, 2023

Hot Coomassie Staining
General protein staining with Coomassie R-350 in hot acetic acid:This hot Coomassie-staining is staining and fixing simultanuously! Works also with native gels, not with IEF-gels. The treatment with Trichloroacetic acid is not necessary.
                       Sensitivity should be: 10 ng per band
The acetic acid for staining and the Ethanol for destaining can be of technical quality.
The staining tray with a metall net for holding the gel away from the magnetic bar:

Because there is no alcohol compound in the staining recipe, this Hot Coomassie staining will also stain membrane-proteins.
During normal, alcohol containing staining recipes, the membrane-proteins get soluble again even after TCA-fixing. These proteins will be not anymore present in the gel!

The assembling of the Hot Coomassie Staining in a steel tray: The staining tray is placed on a magnetic heater. The gel lays gel-face down on the grid. Below the grid stirs the magnetic bar


Staining Tray normal size

Stock solutions
Staining solution:  0.03% (w/v) Coomassie R-350 (
Sigma B4921-20TAB*)
1 tablet (corresponds to 0.4 g dye substance) in 1,25 l 12.5% (v/v) acetic acid (HAc)

Sensitivity enhancement (in progress at EDC):
Addition of 0.01% (w/v) Coomassie Violet R200 (Acid Violet; CI 42650, Sigma #210579-50g) + 0.00065% Ponceau S
Example: 2 tablets R350 + 250 ml HAc + 2.250 l Bidest + 250 mg Coomassie Violet + 16 mg Ponceau S
---> staining 45 min; destaining #1: overnight, destaining #2 & 3: 2 x 60 min, preserving: 30 min

Destaining solution:  12.5% (v/v) HAc
Nicest destaining (with no loss of lipophylic compounds) is: 12.5% HAc overnight, then 2 x 1 h
Preserving solution:  5%-10%* (v/v) Glycerol (*depends on %T of the gel and the air-humidity

EDC’s staining cabinet:

Staining programmes:

[Hot-Coomassie 1D]
[Hot-Coomassie 2D]

 * 0,4 g dye per tablet


EDC Electrophoresis Development & Consulting, Vor dem Kreuzberg 17, 72070 Tübingen (Germany)