Hello everyone. I remember the screen was again from Jenabioscience and this had happened with one of my protein. The screen was very old and the condition was peg3350, tris pH 8, lithium sulfate and NaCl as the salt. Hit was obtained which was never reproducible. Luckily I solved the structure from the hit itself which diffracted to 2.2A resolution. But it is still a mystery for us but this is all crystallography is. Strange things happen.
Faisal Postdoc PHRI, NJ, USA On Jul 31, 2017 5:19 PM, "Janet Newman" <janet.new...@csiro.au> wrote: > Hi Jonathan, > > > Hopefully you know about the trick of making any precious condition last > longer - use the 'magic' solution only in the drop itself, and use the best > approximation you can make in the reservoir of the experiment (if you are > doing vapour diffusion) > > > Regards, Janet > > > Janet Newman > Principal Scientist / Director, Collaborative Crystallisation Centre (C3) > CSIRO Material Science and Engineering > 343 Royal Parade > Parkville. VIC. 3052 > Australia > Tel +613 9662 7326 <+61%203%209662%207326> > Email janet.new...@csiro.au > ------------------------------ > *From:* CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Jonathan > Bailey <baile...@tcd.ie> > *Sent:* 31 July 2017 22:34 > *To:* CCP4BB@JISCMAIL.AC.UK > *Subject:* [ccp4bb] Help needed finding hit condition > > > Dear CCP4bb community > > > I apologies for the slightly off topic post. > > > We have recently had success crystallizing a membrane protein (diffraction > > 3 Å at a synchrotron source) using the *in meso* method, the hit > condition was from the Jena Bioscience screen Pi-minimal condition number > #57. > > > Hit condition – 47.1 % w/v PEG1000, 150 mM Tris pH 8.0, 80 mM Potassium > bromide > > > The screen is old and expired 12/20/2013 (lot # JBS00013133), we have > tried to reproduce the crystals using homemade optimization screens around > the hit condition but have not had any success. We have tried reproducing > the hit using a new (not expired) Pi-minimal screen but had no success. We > are only able to reproduce the crystals using the expired screen and we do > not have much of it left. > > > > We went back and tested the pH of the condition that had given crystals, > the expected pH was 7.9 but we found it to be 6 – 6.5 using a pH indicator > strip. We believe the drop in pH is caused by oxidative degradation of the > PEG1000 resulting in the formation of carboxylic acid species. > > > We have contacted Jena Bioscience to try and get some of the old screen > stock but unfortunately they do not have any. > > > My question is does anyone out there happen to have any expired screen > stocks of this Pi-minimal condition (#57), ideally from the same lot (lot # > JB200013133), that they would be willing to send us. > > > > Does anyone have any advice as to how to reproduce the condition? We’ve > considered bubbling oxygen through and heating the sample to accelerate the > oxidation process. > > > > King Regards > > > Jonathan Bailey (PhD student) > > > Professor Martin Caffrey Lab MS&FB group Trinity College Dublin >
