Re: [ccp4bb] looking for paper describing optimisation of crystals using a screen kit as additive

Sebastiano, I have had really nice success by mixing initial crystallization hits back in to sparse matrix screen conditions. My personal favourite is using the Wizard screens. http://www.rigakureagents.com/p-1-wizard-classic-crystallization-screen-series.aspx I found 75% of the original

Re: [ccp4bb] looking for paper describing optimisation of crystals using a screen kit as additive

Hi Sebastian, you're thinking about local sparse matrix screening. I've done this at a 90:10 ratio. Majeed, S., Ofek, G., Belachew, A., Huang, C.C., Zhou, T., and Kwong, P.D. Enhancing protein crystallization through precipitant synergy. Structure. 2003; 11: 1061–1070 All best. Andreas On

[ccp4bb] looking for paper describing optimisation of crystals using a screen kit as additive

Dear all, I recall a paper (or was is a talk at a conference?) describing the optimisation of initial hits of crystallisation by using a standard screen kit as additive. Something like setting the tray using the initial crystallisation hit condition in the reservoir and mixing 75% of the hit co

Re: [ccp4bb] BSA as additive

Regarding that paper, I would point out that cytosols generally contain 50-100 mM glutamate, so it makes sense that glutamate enhances solubility. JPK From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Evans, Nicola Sent: Monday, May 08, 2017 8:54 AM To: CCP4BB@JISCMAIL.AC.U

Re: [ccp4bb] BSA as additive

I haven't used BSA, but I did recently use 50mM L-glutamic acid for this exact reason (and 5% glycerol in all buffers except the last one for crystallography) after reading this paper and it made a big difference to my last protein prep: https://www.ncbi.nlm.nih.gov/pubmed/15264823 For my fina

Re: [ccp4bb] BSA as additive

(Sorry... over-enthusiastic sending) As for an actual reference - I'm not sure. It's a bit like lab folk-lore. Dave Dr David C Briggs Hohenester Lab Department of Life Sciences Imperial College London UK http://about.me/david_briggs From: CCP4 bulletin

Re: [ccp4bb] BSA as additive

Hello, I've seen BSA used to "block" membranes of centrifugal concentrators when proteins were sticking to them. It appeared to work (proteins could be concentrated with less loss), but I'm not sure how much BSA ends up in your sample. I can imagine the answer is "a bit" and therefore for s

[ccp4bb] BSA as additive

Dear CCP4BB members, Sorry about my non crystallography question. Does anyone know of a reference where BSA has been used as an "additive" in the protein concentration step to prevent aggregation of the main protein? I would appreciate your suggestions. Thank you, H.Sin

Re: [ccp4bb] how to run shelx C/D/E using CCP4 GUI on windows

I cant say anything about Windowssystems - jobs are MEANT to run on either. But re Intensities - you will have intensities in an mtz file somewhere - all data processing outputs both amplitudes and intensities.. As a rule the conversion for I to F and back does not change the stronger intensities

[ccp4bb] how to run shelx C/D/E using CCP4 GUI on windows

Hi everyone, Can anybody tell how to run shelx C/D/E within CCP4 GUI on windows system? Moreover, since shelx C/D/E within CCP4 using mtz file (structure factor) instead of sca file (intensity), would this matters in tough conditions? Thank you! Best regards Chen -- Cheng Chen, Ph.D. Candid

[ccp4bb] Postdoc (Structural Cell Biology) on targeting E3 ubiquitin ligases at Dundee

Job title: Postdoctoral Research Assistant (E3 ubiquitin ligases) Job location: Dundee, UK Job closes: 21-May-2017 Professor Alessio Ciulli’s laboratory is seeking a postdoctoral researcher to undertake research in collaboration with one of the leading pharmaceutical companies supporting the Div