EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Colloidal Coomassie G250 Staining

Mai 30, 2018

All 2DGels can be stained very sensitive and quantitative.
(Attention: use the normal film supported gels, fluor films will not stay more than 30% ethanol!)


Stock solutions:

A) Coomassie Brillant Blue G250 Soluting (CBB-stock)
(5 g Coomassie Brillant Blue G-250, ad. 100mL with Bidest)

B) Colloidal Coomassie Staining Solution (CCD-stock)
(50 g (NH4)2SO4 + 6 ml 85% (w/v) Phosphoric acid, ad. 490mL with Bidest ad 10ml CBB-stock)

C) The final Colloidal Coomassie Solution (CCS-solution)
(200ml CCD-stock+ 50ml MeOH)
Prepare the Colloidal Coomassie Solution alwasy freshly!


Protocol:

2 h fixing in 40% ethanol, 10% Acedic acid
2 x 10 min washing in Bidest
1 day (15 h) staining in Colloidal Coomassie Solution (CCS)
2 x 15 min washing #1 in 20% methanol
2 x 1 h washing #2 with Bidest

ColloidalCoomassieKlein

Colloidal staining in Hoefer’s Autostainer:
The gel should be hold flat by using the 8 magnetic bars
(50% EtOH!)

SDSGel DALT 12.5% Colloidal stained

2D-gel 12.5%T, colloidal stained with Coomassie G250