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Electrophoresis
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Dr. Hanspeter Schickle
Wolfgang Gstrein

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Trouble Shooting Native Electrophoresis

Mai 30, 2018

Gel is leaving its support film:
Do not use staining recipes with alcohol concentrations higher than 40%.
The gel will shrink or swell to much so the forces between the film and the gel get to high!

Troubles-MeOH

No Bands visible
a) Proteins are amphoteric molecules and may run in the reverse direction. Try the opposite native electrophoresis
b) Not every protein is water-soluble at every pH-value. Try the use of soft detergents (Maltosidedodecylsulfate, Triton etc.) +/- 3 M Urea. Check also the opposite native electrophoresis method.
c) Membrane-proteins and other very lipophilic proteins will leave the gel during the staining procedure when alcoholic compounds were present.
Remedy: Use the Hot Coomassie Staining recipe.

Bands smearing. See 1 b)

TSMembraneProtKlein

Curved Electrophoretic Run
hen native electrophoresis is performed very often a enzymatic staining is done. Then the bands can occur low intensiv, although the protein load is too much: The edge of a series of overloaded sample show a curved shape like a waiste.
Remedy: Dilute the samples with the gel buffer

TS-Overload02