In one case of protein crystallization change of buffer ( final purification buffer) gave me better and bigger crystals, but in a different condition.
best Puneet On Thu, Dec 5, 2013 at 6:38 PM, Mahesh Lingaraju <[email protected]> wrote: > 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. >> -- >> > >
