Dear Saurabh I would like to share something that worked for me. I was in a similar situation, though my crystals weren't as beautiful as yours are. I followed Alexander McPherson's Crystallization of Biological Macromolecules and what ultimately worked is as follows: I did something called iterative seeding. That is, I took seeds from generation 1 and grew generation 2, then took seed from generation 2 to grow 3, and so on. It took me about 7-8 generations to get a crystal that was diffracting in the 3 angstrom range. I tried more generations, but the quality didn't really improve anymore. Afterward, I took these crystals and tried to desiccate them before data collection. The method that best worked for me was to dip it in a cryosolution with 5% glycerol --> keep it for 2 mins and then move to 10% glycerol and so on, up to 35% glycerol. You may want to standardize the cryosolution as well. I was able to get up to 2.6 angstrom data at home source.
Hope this helps. *Regards* *Adarsh Kumar, PhD* *Suo Lab* *Florida State University College of Medicine* *Tallahassee, FL - 32304* On Sun, Mar 21, 2021 at 2:04 PM Saurabh Upadhyay <saurabh807...@gmail.com> wrote: > Dear All, > > I am trying to crystalize a protein, which occurs in monomeric (60kDa) and > tetrameric (240kDa) form. *The protein during crystallization was in > MES buffer pH-6 *and the method of crystallization was "*Sitting drop*" > in "*Proplex*" condition. Some of the conditions in which crystals were > obtained are: > > *1) 0.1M HEPES pH-7; 10% (w/v) PEG 4000* > *2) 0.1M Tris, pH-8; 8% (w/v) PEG 8000* > *3) 0.1M Tris, pH-8; 25% (v/v) PEG 400* > > For reference, I have attached the images of the crystals observed below. > > *However, the diffraction pattern obtained and the resolution of crystals > were very poor. Even some mosaicity has been observed. * > > Kindly suggest some methods or modifications, to improve the resolution > and diffraction pattern of the crystals obtained. > > Thanking You, > Sincerely, > Saurabh Upadhyay, > Ph.D. Scholar > c/o Dr. Ashok kumar Patel, > Kusuma School of Biological Sciences, > Indian Institute of Technology, Hauz Khas, > New Delhi-110016, India > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
