Hi Almu, Maybe it will be worth to try even with concentration of 2.5 M salt (or around!) as cryo protectant. Once I picked up crystals directly from drop containing only 2.1M DL-Malic acid and it worked! The protein buffer contained 300 mM NaCl.
To add some more information, crystals were grown in sitting drop with 400 nl total volume and equilibrated against 45 ul reservoir. The crystals were picked up for freezing after around 10 days. Best regards, Mahmudul Lund Sweden On Tue, Oct 30, 2018 at 4:26 PM Mooers, Blaine H.M. (HSC) < blaine-moo...@ouhsc.edu> wrote: > Hi Almu, > > In my hands, 2M LiSO4 is not enough. > > If your crystals cannot survive transfer to higher concentrations, > transfer them to 1.8 M NaMalonate at the same pH and then transfer or add > aliquots of Na Malonate stock at higher concentration until the drop has > about 3.5 - 3.8 M Na Malonate. The optimal final concentration may vary > from system to system. This protocol has worked for us many times. > > Best regards, > > Blaine > > Blaine Mooers, Ph.D. > Associate Professor > Department of Biochemistry and Molecular Biology > College of Medicine > University of Oklahoma Health Sciences Center > S.L. Young Biomedical Research Center (BRC) Rm. 466 > 975 NE 10th Street, BRC 466 > Oklahoma City, OK 73104-5419 > > office: (405) 271-8300 lab: (405) 271-8313 > e-mail: blaine-moo...@ouhsc.edu <mailto:blaine-moo...@ouhsc.edu> > alternate e-mail: bmooe...@gmail.com <mailto:bmooe...@gmail.com> > > Faculty webpage: https://tinyurl.com/BMBmooers > Google Scholar: https://tinyurl.com/GSbmooers > SSRL User Executive Committee: > https://www-ssrl.slac.stanford.edu/content/about-ssrl/advisory-panels/ssrl-users-organization > SSRL-LCLS User Meeting: https://conf.slac.stanford.edu/ssrl-lcls-2018/ > EasyPyMOL: https://github.com/MooersLab > Molecular Graphics Course Links: https://tinyurl.com/molgr > Small Angle Scattering Links: https://tinyurl.com/ouhscsaxs > X-ray lab (LBSF): https://tinyurl.com/ouhsclbsf > ------------------------------ > *From:* CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Almudena > Ponce Salvatierra [maps.fa...@gmail.com] > *Sent:* Tuesday, October 30, 2018 6:44 AM > *To:* CCP4BB@JISCMAIL.AC.UK > *Subject:* [EXTERNAL] [ccp4bb] cryo suggestions for lithium sulphate xt > condition containing 1.8 - 2 M Li2SO4 > > Dear all, > > I have looked into the archive and I've tried to collect suggestions for > cryo protecting crystals that grew in 1.8 M Lithium sulphate containing > mother liquor or 2M. > > What would make more sense? Transferring it into a cryo solution that > consists of mother liquor and a higher concentration of Lithium sulphate? > > Or, would this concentration of Lithium sulphate be enough as a > cryo-protectant and after slight dehydration I could flash freeze them? > > Thank you very much in advance. > > Best > > Almu > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > <https://urldefense.proofpoint.com/v2/url?u=https-3A__www.jiscmail.ac.uk_cgi-2Dbin_webadmin-3FSUBED1-3DCCP4BB-26A-3D1&d=DwMFaQ&c=VjzId-SM5S6aVB_cCGQ0d3uo9UfKByQ3sI6Audoy6dY&r=k0gMbcsdOcdbPUNV5tW66KQSZfXL0ewVDPVBp7tqbks&m=UhcylWwlcufpcF4Jiv9Dy7IwlIWVNuQ1wP5I8uUFr04&s=d92INOuMfDBtY9c9rFVaVpHozWUaoRA9ha6coy-umcI&e=> > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
