depending on how extensively you have screened so far, the most efficient thing to do may be to change the protein: different orthologs, truncations, mutagenesis of "entropy rich" clusters, change of tag location or tag cleavage etc.
________________________________ From: CCP4 bulletin board [[email protected]] on behalf of Mahesh Lingaraju [[email protected]] Sent: Thursday, December 05, 2013 5:38 PM To: [email protected] Subject: Re: [ccp4bb] small crystals Hi All On similar lines, I have been trying to optimize crystallization conditions for my protein. Initially, I had showers of needles in a PEG screen which did not really improve after screening around the condition. So, I seeded these needles into all the screens that I have available and I have plate like crystals which do not diffract at home in MPD (~15%), 0.1 M sodium acetate, 0.2 Mgcl2/cacl2 at 294 K. I tried incubating at 287 K, but that did not yield any useful results.The protein itself is in 50mM MOPS, 10% glycerol pH 7.5. I could try to take of glycerol but i cannot concentrate the protein more than ~ 5mg/ml which clearly was not sufficient to achieve crystallization. Any advice is deeply appreciated. Thank you cheers Mahesh On Thu, Dec 5, 2013 at 10:03 AM, mesters <[email protected]<mailto:[email protected]>> wrote: Hi, can you give a bit more information... Can you concentrate the protein easily to a higher concentration, let's say 2-3 times from what you have now, without precipitation? What is the buffer of your protein stock solution at the moment? At what temperature and what precipitant are you using? - Jeroen - showers of crystals -------- Original message -------- Subject:[SURESPAM] [ccp4bb] small crystals From:Careina Edgooms <[email protected]<mailto:[email protected]>> To:[email protected]<mailto:[email protected]> Cc: Hi all Any advice on how to get bigger crystals from conditions that give showers of tiny crystals? I am getting small pretty looking individual crystals but they are too small and they don't seem to grow. In fact, in some instances if left for a couple of days they actually dissolve. I have fiddled around with mother liquor volume, protein concentration as well as drop volume (I am using hanging drop method) but none seem to make any difference and I always get the same tiny crystals. I think I might try microseeding but I haven't tried that yet. Any suggestions or tricks would be welcome Careina. TotalCare Message Security: Check Authenticity<http://www.exchangedefender.com/verify.asp?id=rB5EYXWs004083&[email protected]> -- Dr. Jeroen R. Mesters Deputy and Group Leader Institute of Biochemistry, University of Lübeck Ratzeburger Allee 160, 23538 Lübeck, Germany phone: +49-451-5004065<tel:%2B49-451-5004065> (secretariate 5004061) fax: +49-451-5004068<tel:%2B49-451-5004068> http://www.biochem.uni-luebeck.de<Http://www.biochem.uni-luebeck.de> http://www.iobcr.org<Http://www.iobcr.org> [X] -- If you can look into the seeds of time and tell which grain will grow and which will not, speak then to me who neither beg nor fear (Shakespeare's Macbeth, Act I, Scene 3) -- Disclaimer * This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. * E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. If verification is required please request a hard-copy version. Please send us by fax any message containing deadlines as incoming e-mails are not screened for response deadlines. * Employees of the Institute are expressly required not to make defamatory statements and not to infringe or authorize any infringement of copyright or any other legal right by email communications. Any such communication is contrary to Institute policy and outside the scope of the employment of the individual concerned. The Institute will not accept any liability in respect of such communication, and the employee responsible will be personally liable for any damages or other liability arising. Employees who receive such an email must notify their supervisor immediately. --
