EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Hot Coomassie Staining 2D

Mai 30, 2018

The first two washing steps have to be performed when Pharmalytes or Biolytes were present in the first dimension.
Servalytes or Sinulyte in the first dimension: gel goes directly in the hot staining solution!

solution (see hot coomassie)

0.4-0.7 mm

Volume
normal gel*

0.8-1.0 mm

temp

40% EtOH / 10% HAc

2 h - overnight

200 ml

3 h - overnight

ambient

destaining solution

20 min

200 ml

30 min

ambient

fresh staining solution at 50-60 oC
(see picture, fume cupboard!)

30 min

600 ml

60 min

50-60 oC

destaining solution in a tray
(rocking platform)

overnight* + 2 x 30 min

200 ml

overnight* + 2 x 60 min

ambient

preserving solution
(in a tray)

20 min

200 ml

30 min

ambient

Air-drying: Overnight, gel should stand upright.
Abbrev.: EtOH = Ethanol, HAc = Acetic Acid
* overnight in the first destaining solution is optimal!
* large size gels: double volumes

2DOpt#ThioUrea#1AS2Klein

DryGel Kit 2D normal size, hot coomassie-stained (+Coomassie Violet)