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 03, 2025
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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