Glycerol is known to be able to reduce nucleation. This might be countered
by an increase in protein concentration.
Vera, L., Czarny, B., Georgiadis, D., Dive, V., Stura, E.A. (2011)
Practical Use of Glycerol in Protein Crystallization. Cryst. Growth & Des.
11: 2755–2762.
Enrico.
On Tue, 03 Apr 2012 13:49:50 +0200, Toby Longwood
<[email protected]> wrote:
Dear all,
My question is related to a sample preparation.
I’m working with a complex that can be stabilized with glycerol (at least
10%) during purification. The use of detergents does not help. After
purification, the sample is homogeneous (EM) and can be concentrated
(3-4mg.mL-1) . I already set up many drops, changing several conditions
(pH, salt...) but nothing conclusive appeared.
I know that crystallogenesis in presence of glycerol works (Sousa, Acta
Cryst (1995), ...) however, because of the aspect of the drops
(precipitates that seem close to the nucleation phase), I suspect that
the
glycerol can be one of the limiting factors of the protocol.
Has anybody else been already confronted to the same problem? Does
someone
know if there is an alternate additive to glycerol?
Thanks in advance for suggestions/help
With best wishes
Toby
--
Enrico A. Stura D.Phil. (Oxon) , Tel: 33 (0)1 69 08 4302 Office
Room 19, Bat.152, Tel: 33 (0)1 69 08 9449 Lab
LTMB, SIMOPRO, IBiTec-S, CE Saclay, 91191 Gif-sur-Yvette, FRANCE
http://www-dsv.cea.fr/en/institutes/institute-of-biology-and-technology-saclay-ibitec-s/unites-de-recherche/department-of-molecular-engineering-of-proteins-simopro/molecular-toxinology-and-biotechnology-laboratory-ltmb/crystallogenesis-e.-stura
http://www.chem.gla.ac.uk/protein/mirror/stura/index2.html
e-mail: [email protected] Fax: 33 (0)1 69 08 90 71
