Hi, to me it looks like 2 things are happening in the drops, to many nuclei and on these initial nuclei new crystals appear. This may indicate that your protein is not that soluble to start with and that there may be some protein aggregation going on in your protein stock solution. Diluting the protein will not help if the protein stock solute (pH, buffer compound, salt ions, etc.) is wrong and your protein is therefore not happy in solution…. DLS can give valuable insights into the aggregation state of the protein stock.
I would re-purify the protein in a different starting solute and try not to pool to many column fractions, in fact, try several of the main column fraction separately. Good luck, Jeroen __ Dr. math. et dis. nat. Jeroen R. Mesters Biological Safety Officer (BBS) Deputy, Lecturer, Program Coordinator Infection Biology Visiting Professorship in Biophysics - University South Bohemia University of Lübeck Center for Structural and Cell Biology in Medicine Institute of Biochemistry Ratzeburger Allee 160 23562 Lübeck Tel +49 451 3101 3105 https://orcid.org/0000-0001-8532-6699 Am 06.12.2024 um 03:18 schrieb 白雪慧 <zb20193020...@cau.edu.cn>: 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 <图片1.jpg> ######################################################################## 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/
