Vitali, Echoing what Dan said, I am not sure why you have chosen detergents first, as there are many other agents which stabilize proteins. Is the instability due to hydrophobic surfaces (e.g., made worse at higher salt) or not. Some non-detergent suggestions are:
1) diols like MPD (see work from Gil Prive's group) 2) non-detergent sulfobetaines (NDSBs), which is the head group in the zwittergent class of deterents. 3) Trimethylamine oxide, which is the head group in the amine oxide class of deterents (LDAO). 4) 200-500 mM L-Arginine 5) Also try 200-500 mM LiCl. The recommendations you received for detergents are very good ones, but remember that many of these detergents are quite dirty, as well as being chemically heterogeneous. Tween 80, Tween 20 and Nonidet P-40 are generally sold at industrial purity (i.e., they are delivered in tanker cars), so either clean them up yourself or buy purified samples (like the Pierce Surfact-Amps brand). Also, be aware that they may be hard to get rid of when you come to setting up crystals. Also look up Sam Gellman's work on detergent-assisted refolding for other stabilizing detergents. Good luck, Michael **************************************************************** R. Michael Garavito, Ph.D. Professor of Biochemistry & Molecular Biology 603 Wilson Rd., Rm. 513 Michigan State University East Lansing, MI 48824-1319 Office: (517) 355-9724 Lab: (517) 353-9125 FAX: (517) 353-9334 Email: [email protected] **************************************************************** On Oct 12, 2012, at 12:42 PM, Vitali Stanevich wrote: > Hi, > > Sorry for off-topic question. > > Does anyone have experience of the stabilisation of water-soluble proteins by > detergents? Protein I'm working with is definitely water-soluble and has high > yield, but, unfortunately, not very stable. Especially during concentration. > So, we thought that adding some detergents may one of the ways to stabilise > protein. > > So, did anyone do it before or may be know published examples? Any > suggestions on the detergent type/concentration would be welcome. > > Thanks, > Vitali
