Dear Dhiraj, the 3C protease is from the Cysteine protease family. Therefore for sure, you can add EDTA (if it is not indented for follow up IMAC purification) and that will inhibit most of the metalloprotease. Furthermore, you can resort to Serine type protease inhibitors like PMSF or Aprotinin.
One side not, the 3C cleavage does not need to be long, as it is an extremely potent protease. If the cleavage sequence is well accessible in less than 1 hour at 4*C you can obtain >95% cleavage when you use 1:100 ratio protease to substrate. Increasing the temperature speeds up the cleavage dramatically and at room temperature, you can obtain the same efficiency for 10 mins approx. I am assuming you are interested in using the 3C protease to cleave a tag from recombinantly expressed protein. Kind regards, Nikolay > On 02/03/2022 6:25 PM Srivastava, Dhiraj <dhiraj-srivast...@uiowa.edu> > wrote: > > > Hi > sorry for off topic question. does anyone know which protease > inhibitors we can include safely while cleaving with 3C protease? > > Thank you > Dhiraj > > > --------------------------------------------- > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > Nikolay Dobrev Postdoctoral Fellow @ Wilmanns group EMBL Hamburg, c/o DESY, Building 25A, Notkestraße 85, 22607 Hamburg, Germany T +49 40 89902 165 | M +49 173 684 0532 twitter.com/emblevents https://twitter.com/emblevents |http://facebook.com/embl.org | http://youtube.com/user/emblmedia Visit http://www.embl.org/events for a complete list of all EMBL events. ######################################################################## 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/
