Dear Everton and Nukri, Thank you for your kind reply, we were also speculating that there could be some issues with cryoprotection.
We have used two cryoprotectants naming; Parabar (10312) and Paraffin Oil from Hampton Research company. We have also used glycerol and respective mother liquors as well. However, there were no improvements in the diffraction pattern or resolution. Can you suggest some modifications regarding cryoprotectants or some ambient temperature for cryoprotection. Kusuma School of Biological Sciences, Indian Institute of Technology, Hauz Khas, New Delhi-110016, India On Sun, 21 Mar 2021, 23:42 Nukri Sanishvili, <[email protected]> wrote: > Dear Saurab, > It's a shame these crystals do not diffract well - they look so good! > I assume the diffraction you saw was recorded at cryo temperatures? If so, > it is possible that you may be damaging the crystals during cryo-pretection > and cryo-cooling. Before you go too deep into changing the crystallization > conditions, I would suggest taking a shot or two at ambient temperatures > and see if the diffraction is any better. If so, you may need to optimize > the cryo protocol and not the crystallization conditions. > Regards, > Nukri > > On Sun, Mar 21, 2021 at 1:04 PM Saurabh Upadhyay <[email protected]> > 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/
