I have purified an intrinsically disordered protein (IDP), and upon running Native PAGE, I observed that it predominantly exists in a dimeric form. I would like to isolate and purify the protein in its monomeric state for further studies. Could anyone suggest effective strategies or conditions (e.g., buffer composition, pH, additives, or purification techniques) that might help in obtaining the monomeric form of an IDP?
If the natural state of this protein is as a homodimer, it would be best to study it in that form.
If it is dimerizing because it is at a higher-than-physiological concentration when purified, it might dissociate into monomers when diluted.
If stabilization of the dimer is mainly by weak hydrophobic interactions between monomers, a mild detergent may shift the equilibrium towards monomers.
If stabilization of the dimer is mainly by ionic interactions, then raising the ionic strength of the buffer by addition of a non-chaotropic salt may shift the equilibrium towards monomers.
Adam B Shapiro My interest of protein is beta synuclein, usually it exists in monomer only. Can you suggest any mild reagent to shift into monomers
Mild, non-denaturing detergents include dodecylmaltoside, n-octylglucoside, CHAPS and others. Non-chaotropic salts include ammonium sulfate and sodium glutamate, for example.
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