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 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/
