Hi Ruben, I have never used TMAO myself but this paper might give you a better idea:
Jeruzalmi, D. & Steitz, T. Use of organic cosmotropic solutes to crystallize flexible proteins: application to T7 RNA polymerase and its complex with the inhibitor T7 lysozyme. J Mol Biol, 1997, 274, 748-56
The authors use 10-20% of cosmotropic solutes. From my experience with several other substances from their list I would use 5-10%.
Best regards, christian Ruben Martinez-Buey wrote:
Hi all! Just want to ask whether some of you has experience with TMAO (or similar conformation stabilizers) for setting up crystallization screenings? Which concentration range would you use? Would you add TMAO to the protein tube and afterwards add the mixture to the plates? Just wanted to know your opinion about conformation stabilizers: could they really help to crystallize multi-domain proteins, supposed to be "flexible"? Thank you very much in advance for your attention, With my best wishes, Ruben
_______________________________________________________________________ Dr. Christian Biertümpfel Laboratory of Molecular Biology NIDDK/National Institutes of Health phone: +1 301 402 4647 9000 Rockville Pike, Bldg. 5, Rm. B1-03 fax: +1 301 496 0201 Bethesda, MD 20892-0580 USA _______________________________________________________________________
