Coommassie Violet

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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			Coomassie-Violet Staining

	Lipohilic proteins will disappear or at least show broader bands during the staining procedure with alcohol containing recipes. As a non-alcoholic procedure we suggest to use the “semi-colloidal Coomassie-Violet” staining. Coomassie-Violet R200 (Acid Violet; CI 42650) Sigma #210579-50g

		The procedure: all steps in ambient temperature!

April 28, 2023

Fixing	20% TCA	200 ml	30 min
Rinsing 1	10% Ethanol	200 ml	20 min
Rinsing 2	H₂O dist.	200 ml	20 min
Staining	0.01% Coomassie-Violet 3,4% (w/v) Phosphoric acid + 3% (v/v) HAc Mix shortly before use!	400 ml	30 - 50 min (staining begins within minutes)
Enhancing (if necessary)	200 ml H₂O + 3 ml dye concentrat (= 0.0015% violet) + 3 ml hot coomassie concentrate (= 0.00045% R350)	200 ml	overnight
Washing + Preserving	5% Ethanol + 5% Glycerin + 5% Ethylenglycol	200 ml	20 min


		Stock solutions: Dye-Concentrate (10 x conc): 0,1% Coom.Violet = 1 g + 1 l H₂O dist Mixing of 400 ml staining solution: To be done shortly before use! ca. 340 ml H₂O dist. + 40 ml dye-conc. + 12 ml HAc + 17 ml H₃PO₄

	Abbrev.: TCA = Trichloric Acide, HAc = Acetic acid

				half IEFGel bidirect 104S stained with Coomassie-Violet incl. Enhancing



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