Thanks everyone,
I got my crystals with PEG 8000 at first and after micro-seeding aith PEG
3350.Now I would work with all your suggestions and references with new
vigor.
ivan
On Thu, Jul 28, 2011 at 9:43 AM, Patrick Shaw Stewart wrote:
> Ed (and Ivan)
>
> Peter Sun and colleagues published two
Ed (and Ivan)
Peter Sun and colleagues published two papers where they show that
crystallization conditions for protein-protein complexes are strongly
biased towards PEG-based rather than high-salt or
organic-solvent-based conditions. This includes antibody-antigen
complexes.
http://www.ncbi.nlm.
On Thu, 2011-07-28 at 05:07 +0100, Sean Seaver wrote:
> Spoiler - Fabs like ammonium sulfate.
Not really - in my hands the ammonium sulfate was one hit out of 7.
While Ivan's question is about Fab complexes with protein antigen, I
think it brings up a more general question of protein class-depend
Hi Ivan,
Here is another example of a method to crystallize antibody/antigene
complexes.
It uses a limited proteolysis step to generate crystals of poor quality,
which are then used as seeds for an MMS screening...
http://www.ncbi.nlm.nih.gov/pubmed/21536542
Good luck,
Alex
2011/7/28 xaravich
Hi Ivan
Did you use microseeding with *random *solutions?
If not see the following paper by Obmolova and Co about exactly this,
microseeding with Fab complexes,
http://journals.iucr.org/d/issues/2010/08/00/bw5361/bw5361.pdf
For more subtle variations in using microseeding with complexes see
http
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
Dear Ivan,
If you take a look at the MPCD ( http://www.cinam.univ-mrs.fr/mpcd/ ) and
search for Fab it brings up 172 crystallization conditions. The names should
help you narrow down, which conditions are of a complex. Hopefully, you can
use those conditions to help you decide, which directio
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 purifi
