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Hi Timur, have you tried seeding from your microstalline stuff? Might be worth to try! Cheers, Albert Albert GUSKOV (Dr) | Research Fellow | Division of Structural & Computational Biology | Nanyang Technological University Proteos 7-01, Biopolis Drive 61, Singapore 138673 Tel: (65) 6586-9690 GMT+8h | Cell: (65) 8366-2779 | Email: [log in to unmask] | Web: www.ntu.edu.sg 2011/7/18 F. Timur Senguen <[log in to unmask]> > Hi everyone, > > I have been issues with a particular protein. I have been close for a > while, but yet so far. > > Rather than going from a clear drop to crystal, my protein first undergoes > phase separation (large oily drops) in which one phase contains most, if not > all, of the protein. This phase separation occurs within a day of preparing > the drop. A day after phase separation the oily phase is now a large > disordered crystalline mass which does not diffract very well. I have tried > changing buffer concentrations, precipitant amounts, ionic strengths and pH > and in all cases this phenomenon is observed. I even screened protein > concentrations to see if reducing protein concentration would prevent the > phase separation. > > Is there any way to bypass this phase separation, which I think prevents me > from obtaining nice crystals. Should I try detergents, chaotropes, or other > additives? > > Thanks in advance. > > Timur > > -- > F. Timur Senguen, Ph.D. > Postdoctoral Research Fellow > Boston Biomedical Research Institute > 64 Grove Street, > Watertown, > MA 02472 USA > >