Hi
I have expressed and purified 16 kDa protein and found this protein forms intermediate of monomer and dimer when it is highly concentrated. From some informations, it might be mediated by hydrophobic residues on its surface.
The sample concentration is 0.5 mM in 20 mM MES pH 6. I have tried additives including 0.2 % Triton X-100, 10 % glycerol, 0.2 % CHAPS and 50 mM Arg. And only 0.2 % CHAPS slightly showed better result.
Any advice or suggestions?
You may not be able to prevent this...
In terms of Le Chatelier’s principle then there is likely to be an equilibrium between monomer and oligomers. The push to the oligomer side is favored by high concentration of monomer. Dilute conditions would favor the mononer state. In some cases an optimum level of ionic stabilier such as phophate may help. Surfactants are only wetting agents (unless ionic themselves) allowing hydrophobic materials to be wetted.
Тry to destroy the hydrophobic bonds by concentrated solutions of urea, guanidinium chloride 4-6 mol / l. The destruction of hydrophobic bonds by means of surfactants requires the selection according to the concentration and the sign of the charge. They can destroy and strengthen hydrophobic bonds. You can try other hydrotropes.
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