[ccp4bb] What's happened over the last five years with high-throughput protein crystallization screening?
Dear CCP4BB Community, I would like to comment you guy a review of the last five years of high-throughput protein crystallization screening that would be a magnificent help for all scientists that struggle with macromolecular crystallization. Please see attachment, as well as below: https://www.tandfonline.com/doi/full/10.1080/17460441.2018.1465924 Thanks. Kind regards Frank Lin To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB=1
Re: [ccp4bb] What's happened over the last five years with high-throughput protein crystallization screening?
Dear Daniel, I appreciate you for your really worthwhile comments that would be useful for other scientists in this field. Thanks. Frank On Thu, Apr 26, 2018, 17:13 Daniel M. Himmel, Ph. D. < danielmhim...@gmail.com> wrote: > I skimmed your paper, and overall it looks like a good overview > of high-throughput protein crystallization. However, I was surprised > that no mention was made of Formulatrix Rock Maker software, > which is an excellent computer-aided graphical tool for designing > crystallization screens rapidly. This software works either as a > standalone > or in conjunction with the Formulatrix Formulator (which the paper DOES > mention), > for preparing crystallization solutions, and/or an integrated system for > storing crystal drops and automatically photographing them under a > microscope. > > A great deal of experience and knowhow on high-throughput > protein crystallization was accumulated by > such researchers as Wladek Minor, Steve Almo, Jeff Bonanno, > and others, with whom I had the privilege of working during the > waning years of the "Protein Structure Initiative" and "Enzyme > Function Initiative". The controversy of these projects stemmed > from the high expense of the robotic equipment that made them > possible, but the methodologies developed and lessons learned > may be useful for high-throughput protein crystallization both > in academia and industry. Hopefully the cost of the robotic equipment > will come down. > > -Daniel > > > On Thu, Apr 26, 2018 at 11:43 AM, Yibin Lin <yyb...@gmail.com> wrote: > >> Dear CCP4BB Community, >> >> I would like to comment you guy a review of the last five years of >> high-throughput protein crystallization screening that would be a >> magnificent help for all scientists that struggle with macromolecular >> crystallization. >> >> Please see attachment, as well as below: >> >> >> https://www.tandfonline.com/doi/full/10.1080/17460441.2018.1465924 >> >> Thanks. >> >> Kind regards, >> >> Frank Lin >> > >
[ccp4bb] crystals disappear
Hi forks, I got some membrane protein crystals in DDM. However, after one month, these crystals change to Irregular shape, and have not birefringent in polarized Light. Do anybody know why? Does the effect of DDM? Contain too much detergent? Thanks, Zhang Peng
[ccp4bb] Crystals
Hi fork, I am working with a membrane protein. I got some crystals from peg400. All of these crystals don't diffract. I tried to use a needle to break crystal. They didn't break into small one, look like gel, or oil, something like that. Do anybody has deal about these crystals. Detergent crystal? My protein contains DDM. Thanks. Frank,
[ccp4bb] Expression of a Selenomethionine Variant in E. coli
Hi List, I want to preprare sel-met labeling protein. Could someone can tell me if it is necessary to add DTT or BME to medium, which contains sel-met, when growing cells? Thanks in advance, Yibin Lin
Re: [ccp4bb] setting up additive screen
Use robot. You only need 0.1ul*96=9.6ul of protein solution. On Thu, Mar 3, 2011 at 7:59 PM, m zhang mzhang...@hotmail.com wrote: Dear all, I am trying to optimize my crystal with additives. Since the yield of my protein purification is very limited, I am wondering what is the most efficient way to set up drops with additive to save my protein and not wasting additives? I am setting up 1 to 1 drops with 0.2 ul additives. But I feel 0.2ul is not very actuate, even if I use a p2. Would you share your ways to set up drops with additives? If I want to screen some additives, what additives would you suggest to try first, especially those from the 96 additive conditions from Hampton? By the way, just wondering, what kind of p2 pipettor work better? Any input is greatly appreciated! Thank you, Min
Re: [ccp4bb] Could someone can help me to explain why EDTA-2Na can formate salt crystals
No, there are not any other cations, so I feel very strange. Everything brought from sigma. On Tue, Feb 22, 2011 at 8:58 PM, William Scott wgsc...@ucsc.edu wrote: What other cations are present? Any divalent cations like Mg++ or Ca++? The Ksp of magnesium phosphate is about 10^-24, so even if you have a very small amount present, say as a contaminant with citrate or EDTA, it will crystallize. On Feb 21, 2011, at 1:22 PM, Yibin Lin wrote: Dear all, I got a lot of salt crystals in reservior solution (well solution), which contains 0.1 M phosphate/citrate ph 4.2, PEG200 47%, EDTA-2Na 0-22mM. Reservior solution appears crystals from 12mM EDTA. Could someone help me to explain why? Thank you very much! Yibin William G. Scott Contact info: http://chemistry.ucsc.edu/~wgscott/
[ccp4bb] Could someone can help me to explain why EDTA-2Na can formate salt crystals
Dear all, I got a lot of salt crystals in reservior solution (well solution), which contains 0.1 M phosphate/citrate ph 4.2, PEG200 47%, EDTA-2Na 0-22mM. Reservior solution appears crystals from 12mM EDTA. Could someone help me to explain why? Thank you very much! Yibin
Re: [ccp4bb] low expression and aggregation of protein
Actually, it is very difficult to say something just based on your simple information. You should give more informations about your work, for example, temperature, cuture, etc. Best, Lin On Fri, Jan 21, 2011 at 8:12 PM, m zhang mzhang...@hotmail.com wrote: Dear All, Recently I am trying to express a glycoprotein in Drosophila S2 cells. However, the expression yield is every low and most of the protein expressed aggregate. I was suggested making new constructs. But I am wondering if anyone here can give me some suggestions to improve it beyond that since I am new to glycoprotein? All your input will be greatly appreciated! Regards, Min
[ccp4bb] selenomethinine-labeled protein
Dear all, I try to express selenomethinine-labeled protein in E.coli methinoine auxotrophic strain B834 in M9 with 17 amino acid and 50 mg/ml se-met for 18 hours at 37 degree. I don't why the colour of medium vary from colorless to yellow. Could anybody tell me it is normal or not? Thanks a lot! Lin
