Not sure if it will be helpful... but my protein is not the most
stable protein, in fact, it does aggregate over time (most likely due
to its 'sticky' nature).
However, I still get crystals. The problem is the crystals are among
the gunks and precipitates.
Your case might be different since my protein does not elute out at
void volume. Perhaps, try work faster? Sometimes protein aggregates
over time rather than immediately.
Allan
Quoting Raji Edayathumangalam <r...@brandeis.edu>:
Hi Folks,
As crazy as it sounds, if you have crystallized and managed to solve the
structure of a protein from aggregated protein, please could you share your
experience.
After many constructs, many many expression schemes and after the usual
rigmarole of optimization that is also often discussed on ccp4bb (buffers,
glycerol, salt concentrations, pH, detergent, additives etc.), I now have a
decently expressing truncated construct for my protein (80 kDa) that is
pure but aggregated (elutes in the void volume from a Superdex200 column).
I am tempted to make a boatload of aggregated protein and set up some
crystal trays (after perhaps testing by CD). So I'd like to hear from folks
who have been successful in solving structures from aggregates when many
many known and tested optimization methods still leave one with aggregated
protein.
Thanks.
Raji
--
Raji Edayathumangalam
Instructor in Neurology, Harvard Medical School
Research Associate, Brigham and Women's Hospital
Visiting Research Scholar, Brandeis University
--
Allan Pang
PhD Student
G35 Joseph Priestley Building
Queen Mary University of London
London
E1 4NS
Phone number: 02078828480
Twitter: @xerophytes
