Hi all,
Recently I have been using Triton X-100 to improve the solubility of my proteins. In order to determine my protein is monomer, dimer, etc, I ran a size exclusion column (S75). Does anyone knows the MW of the Triton X-100 micelle? Another issue is, I still want to test the solubility of my protein so I am gonna remove the Triton X-100. Based on the CMC of Triton X-100 (0.22 mM), I suppose to be able to remove the Triton X-100 by dialysis or Amicorn centrifuge tube after I dilute the Triton X-100 lower than 0.2 mM, am I right? Thanks for you guys' comment.
Shawn
Is your protein tagged? To remove detergent I use to wash resin-tagged proteins with 60% isopropanol, 30% EtOH in general solvents can remove Triton X-100.
To my experience, removal of detergents like Triton by dialysis or washing on a concentrator is very problematic. You can not expect quantitative removal, even at the concentration below CMC. Why don't you use Biobeads SM-2 (BioRad) or Calbiosorb (Calbiochem) resins? Those are polysterene beads with high capacity to lipophilic organics and detergents. You may just add the beads to the protein solution, incubate and remove the beads. Or dialize against a buffer with added beads. There are plenty of publications using this approach. But if you want to remove ALL detergent, then there is no better way than just bind your protein to a resin - Ni-resin, (if tagged), or ion-exchange resins - DEAE- or CM-Sepharose - and wash them extensively. This approach we routinely use in purification of P450's.
The Triton's micelle is like 90 kDa large, so it is not good to use with SEC (my own experience). For that reason we are using CHAPS now.
Also, because of such large micelles, Triton cannot be dialized as micelle, and due to it's small CMC, it will hardly be dialized whenever.
Anyway, my idea was to add some other detergent with smaller micelles, which could wash out the Triton, but I have no idea, whether it would work :)
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