A plea for intellectual assistance...
I am currently purifying a transmembraneous protein from the outer mitochondrial membrane with the aid of the non-ionic detergent Triton X-100. The working concentration of Triton X-100 I am using is 3% (w/v), which is well over the critical micelle concentration (CMC; ~0.033 %) for this detergent. The existence of such micelles aid in the solubilisation of hydrophobic proteins from membrane interiors, but present problems for further downstream chromatographic and enzymatic characterisation.
Does anyone know of a method I could use to disintegrate the micelles formed, without unnecessary detriment to the solubilised proteins?
I have considered diluting the detergent-solubilised protein solution below the CMC, or adding an organic solvent to perhaps demote the thermodynamic favourability for micelle formation and increase the CMC.
If anyone else has any suggestions that might help I would be extremely grateful. Thanks.