The review cited below is a great source of ideas to help you get your ligand in your crystals
Hassell, A et al Acta Crystallogr D Biol Crystallogr. 2007 Jan;63(Pt 1):72-9. Crystallization of protein-ligand complexes. Hopefully it may help you! Dave On 11/12/2007, Schubert, Carsten [PRDUS] <[EMAIL PROTECTED]> wrote: > Simon, > > solubilize your ligand in DMSO so it is maximally concentrated, 100mM works > fine. Add enough compound to achieve 2-3 fold excess to your protein, mix and > set up. Make sure your final DMSO concentration is ~3%, otherwise chances are > you might harm your protein. If you cannot achieve a high enough stock > concentration of DMSO to be below the 3% threshold, dilute you protein in the > storage buffer to ~1mg/ml. Add compound to 2-3 fold access, incubate and > co-concentrate to the desired concentration. That way you avoid the DMSO > shock. > > Alternatively you could incubate the concentrated protein with the compound > solubilized in water for 24-48hr and hope it is soluble and potent enough to > get taken up by the protein and then set up your trays. > > HTH > > Carsten > > > > > -----Original Message----- > > From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] > > Behalf Of Yue Li > > Sent: Tuesday, December 11, 2007 11:55 AM > > To: CCP4BB@JISCMAIL.AC.UK > > Subject: [ccp4bb] insoluble ligand > > > > > > Hi all, > > > > I have one ligand which is insoluble in water, and I would like to > > co-crystallize it with my protein. Is there any other method > > except for > > dissolving it in DMSO ? > > > > Thanks > > > > Simon > > > > > -- ============================ David C. Briggs PhD Father & Crystallographer http://personalpages.manchester.ac.uk/staff/David.C.Briggs/ AIM ID: dbassophile ============================
