- Jürgen Bosch Johns Hopkins Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Phone: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-3655 http://web.mac.com/bosch_lab/
On Aug 28, 2010, at 8:11 AM, 胡小鹏 wrote: > > > 4. Do you have inhibitor in cryo/mounting solution? > > No, I just sock the crystal very quickly (1S) in 30% glycerol. > > Bad idea, even if your ligands are not very soluble have them around in your cryo. Perhaps the solubility is even higher in Glycerol and you are loosing your nicely bound inhibitor even faster. See my previous postin even one second is enough to soak in a ligand so why shouldn't it be enough to loose it in your cryo step. Try optimizing your conditions in such a way that you can directly freeze them out of the drop. Jürgen
