I need help with a bit bizzare (?) problem.
I am trying to overexpress and purify
a small, mostly hydrophobic, most likely
a membrane protein.
Despite correct cloning into pET vector,
I cannot see the overexpressed protein band
after running the total (boiled in SDS) cell
protein after SDS-PAGE.
More troublesome is that the fusion of the
same protein to the maltose-binding protein
(MBP) is sticking to the walls of the plastic
eppendorf tubes after HPLC purification.
Does anyone know what I should do to avoid this,
either to prevent binding to the tubes walls, or
some way to solubilize it without major damage to
the protein structure?
Thanks a lot in advance!
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