EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Lipophilic Proteins

Cause: After the separation and the fixation lipophilic proteins can get native again even after the gel swam 20 minutes in 20% TCA!
This can only happen if alcoholic staining recipes are used. During this procedure the proteins begin to diffuse, the bands get broader and finally the proteins will leave the gel. Hydrophilic proteins are well stained synchronously.
Remedy: will be the use of non-alcoholic staining recipes.
We suggest to use the Coomassie-Violet staining procedure.

To achieve a clear background as Ampholytes only Servalytes, Sinulytes or SepaLytes should be used. Pharmalytes (GE) give strong background!

Coomassie-staining with alcohol
(lipohilic proteins are not visible)

IEFTrouble1003

alcohol-free Coomassie-Violet staining
(lipophilic proteins are visible)

IEFTrouble11

Mai 30, 2018

Coomassie-Violet staining with Pharmalytes
(Pharmalytes-background)

IEFTrouble14