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 patr...@douglas.co.uk
wrote:
Ed (and Ivan)
Peter Sun and
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)
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
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
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
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.
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
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
