Hi Christine Do you know the conditions for crystallization of the Fab only ? You might start using the Pro-complex screens to start with and then use DMSO solubilized peptide in and around that solution.
Thanks, Ivan On Mon, Jul 19, 2010 at 10:37 AM, Harman, Christine < [email protected]> wrote: > Hi all, > I was wondering if anyone has had any experience with setting up > Fab:peptide complexes for crystallization. I have Fab protein and peptide > and I am not sure how to add the peptide to the Fab protein. My peptide is > hydrophobic and thus not very soluble in standard aqueous buffers and barely > soluble in some organics. The Fab protein I have is currently in 100mM > Sodium Acetate pH5, 150mM NaCl. My current plan is to dissolve lyophilized > peptide first in 10% DMSO and use this stock to add to Fab protein in a 20:1 > or 10:1 molar ratio. I am not sure of the effects of residual DMSO on > peptide binding to Fab or on crystallization. I have read one paper in > which they rave about DMSO as an additive in their crystallization, but that > protein was not Fab like at all. Any insight you all might have on this > would be greatly appreciated. > > Thanks in advance, > > Christine > >
