Hi Ivan,
you might also want to find out what buffers your particular system likes.
Jancarik et al. Optimum solubility (OS) screening: an efficient method
to optimize buffer conditions for homogeneity and crystallization of
proteins. Acta Crystallogr D Biol Crystallogr (2004) vol. 60 (Pt 9) pp.
1670-3
Andreas
On 28/07/2011 4:40, xaravich ivan wrote:
Hi everyone,
I have been trying to crystallize Fab:antigen complex( 50kda:90kDa)
complex and initially got needle clusters which after microseeding gave
me single crystals but they are very small and I could not repeat the
results. I have been using HEPES buffer at pH 6.8 to do the final SEC
purification step of the complex before setting trays.
I was wondering whether there are some other buffers (that one could
suggest eg tris-hcl etc) which have given decent positive results when
crystallizing Fab complexes.Though I have gone through individual papers
(case by case) to get some idea, It would be great if anyone could
direct me to a comprehensive literature towards studying the
crystatllization conditions of Fab complexes.
Equally, people who have considerable experience could suggest a list
of must do steps for such problems which have routinely been practiced
in their lab
Also what is a good storage condition for the excess complex that you
want to use later?
I would really appreciate any suggestion,help, direction.
Thanks
ivan
--
Andreas Förster, Research Associate
Paul Freemont & Xiaodong Zhang Labs
Department of Biochemistry, Imperial College London
http://www.msf.bio.ic.ac.uk
