Dear Julie, Thank you for your kindly reply.I do just pool the very central portion of the SEC peak and use that for crystallisation but when SEC Again sill wide peak. And I will try to use other methods like native PAGE to understand in more detail the range of oligomerisation states my protein.
Best Wishes Guohui,SHANG -----原始邮件----- 发件人:"Julie Tucker" <[email protected]> 发送时间:2020-09-15 16:07:00 (星期二) 收件人: [email protected] 抄送: 主题: Re: [ccp4bb] A Problem of my Research Protein Dear Guohui, Sometimes it can work if you just pool the very central portion of the SEC peak and use that for crystallisation? The width of the SEC peak will also depend on factors such as column volume, flow rate, load volume... You do not specify these in your message. Have you tried using other methods such as DLS, DSF, SEC-MALLS, non-reducing SDS- and/or native PAGE to understand in more detail the range of oligomerisation states your protein adopts and their interconvertibility? Best wishes, Julie On Tue, 15 Sep 2020 at 07:22, Guohui SHANG <[email protected]> wrote: Hi Everyone， Well,My Research Protein is easily Dimerzation caused by Disulfide bond for many Cys.And it leads to the SEC peak too wide about 5 mL. Then I Cut N-ter 30 Amino Acids in which 5 spaced Cys,but the SEC Peak is still wide and hard to Crystallize（as the protein is very pure and clean）.Besides, I have tried different Buffer pH ＜ pI or pH ＞ pI，but the SEC peak Do not Work at all. Anyone could offer your kindly ideas,I would thank you very much！ 发自我的iPhone To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 -- Julie Tucker York Biomedical Research Institute Department of Biology and HYMS University of York Currently working remotely and contactable by e-mail only. Please be patient if you do not hear from me promptly. I choose to work flexibly; please only respond to this message when convenient for you to do so. Email disclaimer 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/
