Hi Gloria, I have extensively used SUMO and echoing Artem's sentiment. SUMO has worked well in my hands for most instances. The additional benefit is that, assuming you use His-SUMO-target, following His-tag affinity purification, you can cleave the SUMO tag, followed by reverse IMAC (i.e., collect the flow-thru) to isolate ~≥95% pure target before SEC.
More recently, I only use the SUMO* mutant for *E. coli* and eukaryotic systems, for simplicity's sake. Expression and purification of the protease in-house is easy to do and there are numerous publications on the topic. pET SUMO is how I've typically used the system (usually pET28). Best regards, Nick On Thu, Dec 12, 2024 at 11:37 AM Gloria Borgstahl <gborgst...@gmail.com> wrote: > Hello friends, Yet another kinda off topic question from me. I am looking > into SUMO tags for my protein of interest to help solubility. Has this > worked for you? If so which SUMO and protease do you use? Can the > protease be made in house? So far I am looking at the pET SUMO system. Is > this the best? Thank you in advance. Gloria > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > <https://nam12.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2FWA-JISC.exe%3FSUBED1%3DCCP4BB%26A%3D1&data=05%7C02%7Cndclark2%40g-mail.buffalo.edu%7C9ae68c63d72e4187c82608dd1acb281c%7C96464a8af8ed40b199e25f6b50a20250%7C0%7C0%7C638696182213256137%7CUnknown%7CTWFpbGZsb3d8eyJFbXB0eU1hcGkiOnRydWUsIlYiOiIwLjAuMDAwMCIsIlAiOiJXaW4zMiIsIkFOIjoiTWFpbCIsIldUIjoyfQ%3D%3D%7C0%7C%7C%7C&sdata=RVqTMZTOCrH8cG4SQEWc1QekoYIA0CzXtByAgVpkDUw%3D&reserved=0> > -- Nicholas D. Clark, PhD (He/Him) University at Buffalo Department of Structural Biology Jacob's School of Medicine & Biomedical Sciences 955 Main Street, RM 5130 Buffalo, NY 14203 ######################################################################## 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/
