Dear Raji Running a blue-native gel with lanes in the presence and absence of a reducing agent could prove quite informative. DLS could also return a quick result on the particle distribution in your sample. In that case I would measure samples as fractionated from the superdex200 and compare the measurements after centrifuging the same samples at 100k x g for one hour.
Best regards Savvas On 22 Feb 2012, at 00:21, Raji Edayathumangalam <r...@brandeis.edu> wrote: > 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 > >
