Hi, Did you try using a different column like Superose 6? This column works well to separate large molecular weight proteins including oligomers. Ideally if your solution is not cloudy (coming out of void volume) those are not aggregates those might be oligomers. HTH, Shya
On Tue, Feb 21, 2012 at 6:21 PM, 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 > > >
