EDC Impressum & DSE

Electrophoresis
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Dr. Hanspeter Schickle
Wolfgang Gstrein

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Coomassie Staining Troubles of IEF-Gels

Mai 30, 2018

Two major problems occur:

1.   Blue stripes along the iso-pH-lines occur after staining and destaining
           Cause: The Ampholytes thet are creating the pH-gradients are also
             “Amin-Carbon-Acides” and though strongly stainable with Coomassie. See below.
              Remedy: Usa longer washing steps after the TCA and/or a higher alcohol concentration
              to get rid of it. See this page: bluelines

2.   Lipophylic Proteins were not stained (at all)
             Cause: After the separation and the fixation lipophilic proteins can get native again
             even after the gel swam 20 minutes in 20% TCA!
             Remedy: See this page: Lipoproteins

TS case #1: Stripes along the iso-pH-lines.Normally in the neutral and/or the acidic region (in this case left side)...

TSIEF-Stripes

...a very hard case

TSbasicPeptides2Klein