There are numerous detergents to try, although DDM is usually my standard choice for an initial try.
Other detergents that I usually try in the beginning are: LDAO, b-OG, NG, DM, NGOG, Cymal-5 and Cymal-6. The concentration used for solubilisation of course needs to be varied depending on the detergent used.
Hi
Ronnie's suggestions are excellent.
Can we get more information form you:
Is this an E. coli membrane or yeast? I am wondering why DDM did not work as it typically works for most membrane solubilization. Are you following a protocol from a paper? Are you solubilizing the protein from purified membranes or whole cells? What concentration of DDM is used?
You can also try CHAPS, it has a large CMC and cholesterol-like structure
http://info.agscientific.com/blog/bid/158851/CHAPS-14-FAQs-Protocols-you-must-have
N-Lauryl Sarcosine may be helpful for solubilizing membrane proteins..
with regard to what M Joanne Lemieux wrote above:
how were your results using DDM? Is the "trouble" you have (I guess?) with the detergent, or protein purification per se in your case? If you could give additional information the people here are surely glad to assist :)
Factors like cell disruption, membrane isolation, protein purification, buffer and storage conditions can greatly influence your successful solubilization and need to be checked for every individual protein (sadly)
Triton X-100 could be effective. What was the trouble with DDM?
you have a number of detergent options depending on what you wish to do with the protein down stream for very mild detergents which will maintain activity and folding you can use digitonin or the new synthetic version GDN form anatrace alternatively if you wish to rip everything out of the membrane and you dont care about activity/fold then use something like a FC12. Each detergent has its own benefits and draw backs such as CMC and harshness so a balance has to be sort....
M Joanne Lemieux .. From bacteria and not yeast. And I am trying to solubilize protein from purified cell membrane. Protein is getting solubilized by DDM which is pure however most fraction is remaining in cell membrane. its not that effective by DDM even after increase in its conc.
Paula .. I cannot use Triton X 100 as I am worrying about misfolding
I had a similar problem with a bacterial protein. DDM was not very effective but DM (5 to 10 cmc) worked nicely. Good luck!
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts