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Re: [ccp4bb] co-crystallization |
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- Protein crystallographyMain steps:- Protein purification- Crystallisation Special:- Programs for crystallography- X-ray detectors Basic tutorials:- Chemistry- Protein - Peptide - Amino Acids Xtal community:- CCP4BB |
CCP4bb navigationCCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999Subject: Re: co-crystallization From: Juergen Bosch jbosch {- at -} U {- dot -} WASHINGTON {- dot -} EDU Date: 2008-12-01 The Km changes with your reservoir, so predictions are limited. In general if you have a low Km this is favourable but not a given that your ligand will be found in the electron density map. As a starting point try a molar ratio of >3 of the ligand to your protein and you can go as high as you want (assuming it's soluble and does not cost you an arm and a leg). 50 fold excess has been used in some cases. And remember for your cryo-step to include your ligand in sufficient amounts ! J¨¹rgen On 1 Dec 2008, at 05:31, yangliuqing wrote: > Hello,everyone, > I have a question for cocrystallization, is there some relationship > between Km value and substrate concentration when making > cocrystallization? How can I know the substrate is enough for binding? > Thank you very much! > liuqing > > °ËØÔÓéÀÖ°ü´òÌý£¬MSN×ÊѶËٵݰïÄã棡 Á˽âÏêϸ£¡ - J¨¹rgen Bosch University of Washington Dept. of Biochemistry, K-426 1705 NE Pacific Street Seattle, WA 98195 Box 357742 Phone: +1-206-616-4510 FAX: +1-206-685-7002 Web: http://faculty.washington.edu/jbosch CCP4bb navigationCCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999 |
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