Dear Friduous,
This is also a good paper to look at once you know your crystals diffract at room temperature: https://www.ncbi.nlm.nih.gov/pubmed/10944336 Many of the salts in your condition will be cryo-protective if you increase their concentration to saturated levels. Cheers, Joseph. Joseph S. Brock | PhD Researcher Drew Lab Department of Biochemistry and Biophysics Stockholm University ________________________________ From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of CCP4BB automatic digest system <lists...@jiscmail.ac.uk> Sent: Sunday, 21 October 2018 1:00:06 AM To: CCP4BB@JISCMAIL.AC.UK Subject: CCP4BB Digest - 19 Oct 2018 to 20 Oct 2018 (#2018-267) There are 5 messages totaling 760 lines in this issue. Topics of the day: 1. suggestions for cryoprotectant (4) 2. RapiData at SSRL 2019 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ---------------------------------------------------------------------- Date: Fri, 19 Oct 2018 17:23:49 -0700 From: Christine Gee <chr...@gmail.com> Subject: Re: suggestions for cryoprotectant Dear Fridous Have a look at https://www.ncbi.nlm.nih.gov/pubmed/14646118 [http://www.ncbi.nlm.nih.gov/coreutils/img/pubmed256blue.png]<https://www.ncbi.nlm.nih.gov/pubmed/14646118> Malonate: a versatile cryoprotectant and stabilizing solution for salt-grown macromolecular crystals. - PubMed - NCBI<https://www.ncbi.nlm.nih.gov/pubmed/14646118> www.ncbi.nlm.nih.gov Acta Crystallogr D Biol Crystallogr. 2003 Dec;59(Pt 12):2356-8. Epub 2003 Nov 27. Malonate as a cryoprotectant Or https://www.ncbi.nlm.nih.gov/pubmed/20606259 [http://www.ncbi.nlm.nih.gov/coreutils/img/pubmed256blue.png]<https://www.ncbi.nlm.nih.gov/pubmed/20606259> Cryoprotection properties of salts of organic acids: a case study for a tetragonal crystal of HEW lysozyme. - PubMed - NCBI<https://www.ncbi.nlm.nih.gov/pubmed/20606259> www.ncbi.nlm.nih.gov Acta Crystallogr D Biol Crystallogr. 2010 Jul;66(Pt 7):789-96. doi: 10.1107/S0907444910015416. Epub 2010 Jun 19. Regards Christine On Fri, Oct 19, 2018 at 2:57 PM Firdous Tarique <kahkashantari...@gmail.com> wrote: > Dear members > > I have got beautiful crystal hits in SaltRx screens which are not > diffracting to a good resoultion. All of them are salt based condition and > I am not able to formulate a good cryoprotectant for these crystals. I also > think that in my case the poor resolution is due to a poor cryoprotectant > selection. > > The conditions are as follows: > > 1> 4M Ammonium Acetate 100mM Bis Tris Propane pH 7.0 > 2>0.5M KCN 100mM Tris pH8.5 > 3>1.5M LiSo4 100mM Bris Tris Propane pH 7.0 > 4>4M Sodium Nitrate 100mM Tris pH8.5 > 5>1.5M Sodium Nitrate 100mM Sodium acetate pH 4.6 > > There are few more conditions but so far not able to see good diffraction > with using lower peg and glycerol based cryoprotectants. > > Can anybody suggest me good cryos conditions for salt based > crystallization conditions or anything good for SaltRx crystallization hits. > > Thanks > > Firdous > > ------------------------------ > > 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 ------------------------------ Date: Sat, 20 Oct 2018 02:21:41 +0000 From: "Smith, Clyde" <csm...@slac.stanford.edu> Subject: RapiData at SSRL 2019 Hi everyone, Applications are now open for the 2019 edition of RapiData at SSRL. RapiData is a practical course in macromolecular X-ray diffraction data collection, data processing and structure solution. The aim of the RapiData course is to educate and train young scientists in data collection and processing methods at synchrotron beamlines, using state-of-the-art software and instrumentation. The course will be held at the Stanford Synchrotron Radiation Lightsource (SSRL) located on the SLAC National Accelerator Laboratory campus in Menlo Park, CA, from May 5-10 2019. Course organizers for 2019 are Silvia Russi, Clyde Smith and Jeney Wierman. The course will comprise hands-on experiments at the SSRL beamlines, software tutorials, and lectures on the following topics: Specimen preparation, Data collection strategies, X-ray light sources, X-ray detectors, Data reduction, Structure solution by MAD, SAD and Molecular Replacement, Complementary methods (spectroscopy and small angle scattering) Please visit http://smb.slac.stanford.edu/news/rapidata/rapidata-2019/ and use the links on the "registration" page to apply for the course. The application deadline is December 31 2018 Successful applicants will be notified early in January 2019 and invited to register and book accommodation at the SLAC Guesthouse at that time. A limited amount of travel support funding may be available. RapiData 2019 at SLAC<http://smb.slac.stanford.edu/news/rapidata/rapidata-2019/> smb.slac.stanford.edu RapiData 2019 is a practical course in macromolecular X-ray diffraction data collection, data processing and structure solution. Please direct questions to Silvia Russi (sru...@slac.stanford.edu), Jeney Wierman (jwier...@slac.stanford.edu) or Clyde Smith (csm...@slac.stanford.edu). ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ------------------------------ Date: Sat, 20 Oct 2018 09:01:51 +0200 From: Andreas Förster <docandr...@gmail.com> Subject: Re: suggestions for cryoprotectant Dear Firdous, get some Mitegen sleeves and test your crystals at RT. Before you establish diffraction, there’s no need to agonize about cryo-conditions. Good luck! Andreas On Fri, 19 Oct 2018 at 23:57, Firdous Tarique <kahkashantari...@gmail.com> wrote: > Dear members > > I have got beautiful crystal hits in SaltRx screens which are not > diffracting to a good resoultion. All of them are salt based condition and > I am not able to formulate a good cryoprotectant for these crystals. I also > think that in my case the poor resolution is due to a poor cryoprotectant > selection. > > The conditions are as follows: > > 1> 4M Ammonium Acetate 100mM Bis Tris Propane pH 7.0 > 2>0.5M KCN 100mM Tris pH8.5 > 3>1.5M LiSo4 100mM Bris Tris Propane pH 7.0 > 4>4M Sodium Nitrate 100mM Tris pH8.5 > 5>1.5M Sodium Nitrate 100mM Sodium acetate pH 4.6 > > There are few more conditions but so far not able to see good diffraction > with using lower peg and glycerol based cryoprotectants. > > Can anybody suggest me good cryos conditions for salt based > crystallization conditions or anything good for SaltRx crystallization hits. > > Thanks > > Firdous > > ------------------------------ > > 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 ------------------------------ Date: Sat, 20 Oct 2018 09:59:45 +0200 From: mesters <mest...@biochem.uni-luebeck.de> Subject: Re: suggestions for cryoprotectant Hi, have a look at the following interesting papers https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466044/ [https://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/pmc-logo-share.png]<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466044/> Multicomponent mixtures for cryoprotection and ligand solubilization<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466044/> www.ncbi.nlm.nih.gov Mixed cryoprotectants have been developed for the solubilization of ligands for crystallization of protein–ligand complexes and for crystal soaking. Low affinity lead compounds with poor solubility are problematic for structural studies. Complete ... https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2852300/ [https://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/pmc-logo-share.png]<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2852300/> Progress in rational methods of cryoprotection in macromolecular crystallography<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2852300/> www.ncbi.nlm.nih.gov Cryogenic cooling of macromolecular crystals is commonly used for X-ray data collection both to reduce crystal damage from radiation and to gather functional information by cryogenically trapping intermediates. However, the cooling process can damage ... https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4252874/ [https://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/pmc-logo-share.png]<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4252874/> Efficient Cryoprotection of Macromolecular Crystals using Vapor Diffusion of Volatile Alcohols<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4252874/> www.ncbi.nlm.nih.gov Macromolecular X-ray crystallography, usually done at cryogenic temperature to limit radiation damage, often requires liquid cryoprotective soaking that can be labor intensive and damaging to crystals. Here we describe a method for cryoprotection that ... Good luck J. Am 19.10.18 um 23:57 schrieb Firdous Tarique: > Dear members > > I have got beautiful crystal hits in SaltRx screens which are not > diffracting to a good resoultion. All of them are salt based condition > and I am not able to formulate a good cryoprotectant for these > crystals. I also think that in my case the poor resolution is due to a > poor cryoprotectant selection. > > The conditions are as follows: > > 1> 4M Ammonium Acetate 100mM Bis Tris Propane pH 7.0 > 2>0.5M KCN 100mM Tris pH8.5 > 3>1.5M LiSo4 100mM Bris Tris Propane pH 7.0 > 4>4M Sodium Nitrate 100mM Tris pH8.5 > 5>1.5M Sodium Nitrate 100mM Sodium acetate pH 4.6 > > There are few more conditions but so far not able to see good > diffraction with using lower peg and glycerol based cryoprotectants. > > Can anybody suggest me good cryos conditions for salt based > crystallization conditions or anything good for SaltRx crystallization > hits. > > Thanks > > Firdous > > ------------------------------------------------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > -- Dr.math. et dis. nat. Jeroen R. Mesters Deputy, Senior Researcher & Lecturer Program Coordinator /Infection Biology/ <http://www.uni-luebeck.de/studium/studiengaenge/infection-biology/introduction.html> Introduction: Universität zu Lübeck<http://www.uni-luebeck.de/studium/studiengaenge/infection-biology/introduction.html> www.uni-luebeck.de Universität zu Lübeck. Im Focus das Leben Institute of Biochemistry, University of Lübeck Ratzeburger Allee 160, 23538 Lübeck, Germany phone: +49-451-31013105 (secretariate -31013101) fax: +49-451-31013104 http://jobs.zeit.de/image-upload/logo_10564.jpg [http://jobs.zeit.de/image-upload/logo_10564.jpg] http://www.biochem.uni-luebeck.de <Http://www.biochem.uni-luebeck.de> http://www.eine-stadt-sieht-gelb.de <Http://www.eine-stadt-sieht-gelb.de> http://www.uni-luebeck.de/studium/studiengaenge/infection-biology Infection Biology: Universität zu Lübeck<http://www.uni-luebeck.de/studium/studiengaenge/infection-biology> www.uni-luebeck.de Universität zu Lübeck. Im Focus das Leben http://www.iobcr.org <Http://www.iobcr.org> Visiting Professorship in Biophysics, University of South Bohemia (CZ) President of the International Organization for Biological Crystallization (IOBCr) -- 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) -- "Aujourd'hui je sais qu'il n'y a pas de limites à la bêtise humaine - qu'elle est infinie." (Gustave Flaubert, French novelist, 1821-1880) -- It is invariably the case that high resolution X-ray structures show significantly better agreement with solution observables such as coupling constants, 13C chemical shifts, and proton chemical shifts, than the corresponding NMR structures, including the very best ones. Hence, in most cases, a high-resolution crystal structure (< 2.0 Å)will provide a better description of the structure in solution than the corresponding NMR structure (Kuszewski, Gronenborn & Clore, 1996, Protein Science 5:1067-80) -- 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. 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Employees who receive such an email must notify their supervisor immediately. -- ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ------------------------------ Date: Sat, 20 Oct 2018 15:03:41 +0000 From: "Thomas, Leonard M." <lmtho...@ou.edu> Subject: Re: suggestions for cryoprotectant This is the first thing you need to do before going down the rabbit hole looking for a cryo protectant. Mitegen make it much easier then in the 'old days' of capillaries. Good luck, Len lmtho...@ou.edu On Oct 20, 2018 2:02 AM, Andreas Förster <docandr...@gmail.com> wrote: Dear Firdous, get some Mitegen sleeves and test your crystals at RT. Before you establish diffraction, there’s no need to agonize about cryo-conditions. Good luck! Andreas On Fri, 19 Oct 2018 at 23:57, Firdous Tarique <kahkashantari...@gmail.com<mailto:kahkashantari...@gmail.com>> wrote: Dear members I have got beautiful crystal hits in SaltRx screens which are not diffracting to a good resoultion. All of them are salt based condition and I am not able to formulate a good cryoprotectant for these crystals. I also think that in my case the poor resolution is due to a poor cryoprotectant selection. The conditions are as follows: 1> 4M Ammonium Acetate 100mM Bis Tris Propane pH 7.0 2>0.5M KCN 100mM Tris pH8.5 3>1.5M LiSo4 100mM Bris Tris Propane pH 7.0 4>4M Sodium Nitrate 100mM Tris pH8.5 5>1.5M Sodium Nitrate 100mM Sodium acetate pH 4.6 There are few more conditions but so far not able to see good diffraction with using lower peg and glycerol based cryoprotectants. Can anybody suggest me good cryos conditions for salt based crystallization conditions or anything good for SaltRx crystallization hits. Thanks Firdous ________________________________ 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=qKdtBuuu6dQK9MsRUVJ2DPXW6oayO8fu4TfEHS8sGNk&r=LYFS_Pt-UDexWUsCj6bDwQ&m=-ToI6ga-FYygb9nKnP_gQXDS5-my58CypHjzkpTAcXc&s=aphz-daUOztDbG1vJsutT1p4M_P9ICmhAl2OkEdU63Q&e=> ________________________________ 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=qKdtBuuu6dQK9MsRUVJ2DPXW6oayO8fu4TfEHS8sGNk&r=LYFS_Pt-UDexWUsCj6bDwQ&m=-ToI6ga-FYygb9nKnP_gQXDS5-my58CypHjzkpTAcXc&s=aphz-daUOztDbG1vJsutT1p4M_P9ICmhAl2OkEdU63Q&e=> ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ------------------------------ End of CCP4BB Digest - 19 Oct 2018 to 20 Oct 2018 (#2018-267) ************************************************************* ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
