Dear James --
One possible issue you might have as well is oxidation. Your sample
might need to be in a different step... and would therefore require
some time before getting to this step. I personally noticed (in few
cases) that when my crystals took time to grow, it was either because
I can't remember if someone has already suggested this. You can
dissolve some of your crystals and ask your favorite mass spec. lab to
check if your protein has been oxidized, proteolyzed, etc.
ho
Confometrx
Dear crystallographers,
Sorry for the non-ccp4 query. I am new to this field and need some
suggestions. My question is, why some protein takes longer time to
crystallize, say 6-8 months, and it is the only condition to get the
crystals.? What are the ways to get the crystals faster.
The crystal
It will help more if we send it to James ;)
On Oct 11, 2009, at 9:15 AM, gauri misra wrote:
Dear James,
As there are indications of protein degradation that have been
suggested in
previous postings, i think adding some protease inhibitors right at
the
stage of purification may provide
Hi James,
have you tried limited proteolysis on your protein and see if you can
identify a stable fragment. Then re-clone and re-crystallize your
protein. Or a very stupid suggestion, how does your size exclusion
peak look like ? What you're not running your protein over a SEC to
polish
Sometimes it is better that it takes time for crystals to appear.
Remember that crystallisation is a purification procedure. A way to
decrease the speed of crystallisation is to use Dunlop's and Haze's drop
dilution method (K. V. Dunlop B. Hazes (2003). When less is more: a
more efficient
