Hi Nicola, Our TurboTEV (http://accelagen.com/TurboTEV-datasheet.htm) is very stable and does not need additional reducing agent for its activity. The storage buffer has 1 mM TCEP. It'll be likely <0.1 mM in your digestion. You can get it from our European distributors or directly from Accealgen. Sigma probably sells it in Europe as well. TurboTEV is shipped with gel packs and can be stored at -20 for long time.
Cheers, Chun Accelagen -----Original Message----- From: CCP4 bulletin board <[email protected]> On Behalf Of Nicola Evans Sent: Wednesday, February 13, 2019 10:46 AM To: [email protected] Subject: [ccp4bb] TEV Protease in low reducing agent? I am interested in purifying proteins with 1 or 2 disulphide bonds, and have been using enterokinase to cleave off N-terminal tags but we have had issues with poor cleavage and want to try TEV cleavage instead. However TEV protease is usually kept in a high amount of DTT and I am concerned about reducing the disulphide bonds in my purified proteins. I have used TEV protease before with 0.25mM TCEP and it worked well. Is there a way to use TEV with very little or no reducing agent? Perhaps by optimising conditions such as adding glycerol? We were thinking of buying in commercial TEV protease so any advice on that is also welcomed (is there any merit, except cost, to making it ourselves?) ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
