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]>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]> > To:[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 (secretariate 5004061) > fax: +49-451-5004068 > > http://www.biochem.uni-luebeck.de <Http://www.biochem.uni-luebeck.de> > http://www.iobcr.org <Http://www.iobcr.org> > > > -- > 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. > -- >
