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Trouble Shooting Native Electrophoresis

April 28, 2023

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!


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)


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


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