Hi ZB I . . . . removed the primary crystal condition
I'm not sure exactly what procedure you are using. You should suspend the crushed crystals in the original crystallization condition and add a small amount (about 15% of the drop) to every drop in a random screen eg Crystal Screen, Index, JCSG+. Your protein seems to crystallize easily, so I will be surprised if you don't get nice single crystals within two or three 96-well screens. my crystals grow within three days If you use seeding, your crystals should grow overnight You need to "let go" of the conditions you're working with - find some new ones! Good luck, Patrick On Fri, Dec 6, 2024 at 2:18 AM 白雪慧 <zb20193020...@cau.edu.cn> wrote: > Thank you very much for your reply to my question on how to optimize > microcrystal. In response to your suggestions and questions, I would like > to make the following reply: > First of all, my protein can grow crystals like this one in a variety of > situations. > For example, other conditions: > 1.15% PEG 8000;0.2M MgCl2;0.1M HEPES pH 7.5 > 2.20% PEG 1500;0.2M MgCl2;0.1M HEPES pH 7.5 > 3.25% PEG 8000;0.2M NaCl;0.1M NaCaco pH 5.5 > 4.20% PEG 1e500;0.2M MgCl2;0.1M Tris pH 8.5 > Second, my crystals grow within three days, and there is no phosphate in > the protein solution. > Finally, in fact, I tried crystal inoculation before, but did not find any > change in crystal shape. Maybe my operation was also wrong. I sucked out > the crystal, mashed it, diluted it in series, and removed the primary > crystal condition, and the protein concentration did not change, nor did > the concentration of the pool liquid change greatly > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > -- patr...@douglas.co.uk Douglas Instruments Ltd. Douglas House, East Garston, Hungerford, Berkshire, RG17 7HD, UK Directors: Patrick Shaw Stewart, Peter Baldock, Stefan Kolek http://www.douglas.co.uk Tel: 44 (0) 148-864-9090 US toll-free 1-877-225-2034 Regd. England 2177994, VAT Reg. GB 480 7371 36 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
