> > It is pretty normal (actually, pretty good) to get crystals within 3 days > under room temperature
Why work with conditions that take 3 days for crystals to grow when you can, very likely, find conditions that grow in one day by using seeding ! ? (I'm simplifying slightly - they may take 3 days to *stop *growing) On Sat, Dec 7, 2024 at 8:00 PM Kevin Jin <kevin...@gmail.com> wrote: > First of all, you need to verify these crystals are protein. To do so, use > a loop to fish some crystals and take a shot to see the reflection pattern, > a lazy way! > > If they are protein, then start considering the optimization. No matter > which kind of kits you have used, they are just matrices of different > conditions. > > It is pretty normal (actually, pretty good) to get crystals within 3 days > under room temperature. The only problem is the xtals grow a little bit > fast, > > Notice your condition, protein solubility is not an issue. The only > problem is the folding force needed to be adjusted. Likely, you need to > decrease the total salt concentration. > > For PEG 8K, you may try 5 ~ 15%, instead of 20~25 %. > > Different polymers, their pH values are different. Based on your protein's > charge, different combinations may give you some unexpected results, > dynamics vs. kinetics process. > > Basically, inorganic chemistry + polymer chemistry/physics + solution > chemistry + protein chemistry > > Good luck with best regards! > > > > > On Thu, Dec 5, 2024 at 6:18 PM 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 >> > > ------------------------------ > > 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/
