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
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