An alternative is to dissolve your compound in MeOH and dispense it either manually or via robot, let the plate sit sometime in the hood for faster evaporation and then add your protein + reservoir. Jürgen ...................... Jürgen Bosch Johns Hopkins University Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Office: +1-410-614-4742<tel:%2B1-410-614-4742> Lab: +1-410-614-4894<tel:%2B1-410-614-4894> Fax: +1-410-955-2926<tel:%2B1-410-955-2926> http://lupo.jhsph.edu
On Oct 15, 2014, at 5:18 PM, Keller, Jacob <kell...@janelia.hhmi.org<mailto:kell...@janelia.hhmi.org>> wrote: Since you mentioned EtOH, why not do this: -Make a tray with the appropriate mother liquors -Make a drop for each well containing mother liquor and high-concentration ligand in EtOH (you could vary the ratios here as needed.) -Equilibrate by vapor diffusion until EtOH all goes into the well soln (couple of hours at the most?) -Add protein to these drops You could skip right to the protein step if your protein doesn't mind EtOH at fairly high concentrations, and anyway it will be gone fairly quickly, esp at RT Jacob Keller ________________________________________ From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK>] on behalf of Monica Mittal [monica.mitta...@gmail.com<mailto:monica.mitta...@gmail.com>] Sent: Wednesday, October 15, 2014 2:13 PM To: CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK> Subject: [ccp4bb] crystallization with hydrophobic ligands Dear All Can anyone give suggestions for handling the solubility problem of highly hydrophobic compounds, during co-crystallization or inhibition assays? The ligands I am using are almost insoluble in aquous medium. In DMSO or 95% Ethanol, the solubility is higher. Besides crystallization, this solubility is also a hindrance for in-vitro or in-vivo assays requiring higher conc. of ligand. Thanks in advance ! Monica
