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
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