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Thanks to everyone for their advice - I was running out of ideas, but once again, hope springs eternal! My apologies for giving insufficient information - lowest was 1-2% PEG in hanging drop. 2% still see microcrystalline ppt in skin. Thanks again, Fi Quoting Tommi Kajander <[EMAIL PROTECTED]>: > hi, check the leaf clumps for diffractio is the most important thing + > microseed at concentration that you dont get microcrystals at (say let sit > for a day?) you dont say what 'lowest' means, go bit lower, also you dont > say _when_ do you get the microcrystals (if they are in the buffer change > buffer eg add salt)? seed with a whisher by poking/crushing a crystal with > it > ans sweep through a couple drops.... > HTH, > t. > > Quoting Fiona Whelan <[EMAIL PROTECTED]>: > > > *** For details on how to be removed from this list visit the *** > > *** CCP4 home page http://www.ccp4.ac.uk *** > > > > > > Hello All, > > I'm attempting to co-crystallize a dimeric transcription factor bound to > > DNA > > and see skin formation and microcrystals at even the lowest concentration > > of > > precipitant that I have tried. The complex crystallizes (I believe) on > > storage > > at 4 deg, but these crystals do not diffract. I have additionally tried > > using > > increased concentrations of reducing agents and also covalent > > modifications of > > Cys residues and still see skin formation. After long term storage, > > larger > > crystals grow after a few months storage, but are leaf shaped and growing > > in > > clumps. I have not checked the diffraction limits of these crystals as > > yet (no > > access to local x-ray source). > > > > Any advice as to how to control the seeding events and/or skin formation > > would > > be appreciated. > > > > Thanks! > > > > Fi > > > > > > > > Fiona Whelan > > Whitelaw Lab > > School of Molecular and Biomedical Science (Biochemistry) > > The University of Adelaide > > Phone: 8303 4806 > > > > > -- > Tommi Kajander, Ph.D. > Macromolecular X-ray Crystallography > Research Program in Structural Biology and Biophysics > Institute of Biotechnology > PO box 65 (Street address: Viikinkaari 1, 4th floor) > University of Helsinki > FIN-00014 Helsinki, Finland > Tel. +358-9-191 58903 > Fax +358-9-191 59940 > Fiona Whelan Whitelaw Lab School of Molecular and Biomedical Science (Biochemistry) The University of Adelaide Phone: 8303 4806 ----- End forwarded message ----- Fiona Whelan Whitelaw Lab School of Molecular and Biomedical Science (Biochemistry) The University of Adelaide Phone: 8303 4806
