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Dear Chun and all, I do have a questions: If I find the patent describing this mutation, (ttps://patents.google.com/patent/CN101864407A/en, maybe it is not), and If I reproduce those mutations in my lab, and If I express and purify these mutated protein and used it in my lab for academic research, does the lab that I represent have to pay anything, as this is a non profit mailing list? looking forward to your answer. Kind regards André ----- Original Message ----- From: "Chun Luo" <[log in to unmask]> To: [log in to unmask] Sent: Wednesday, 13 February, 2019 21:37:27 Subject: Re: [ccp4bb] TEV Protease in low reducing agent? 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 <[log in to unmask]> On Behalf Of Nicola Evans Sent: Wednesday, February 13, 2019 10:46 AM To: [log in to unmask] 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 Helmholtz Zentrum Muenchen Deutsches Forschungszentrum fuer Gesundheit und Umwelt (GmbH) Ingolstaedter Landstr. 1 85764 Neuherberg www.helmholtz-muenchen.de Aufsichtsratsvorsitzende: MinDirig'in Petra Steiner-Hoffmann Stellv.Aufsichtsratsvorsitzender: MinDirig. Dr. Manfred Wolter Geschaeftsfuehrer: Prof. Dr. med. Dr. h.c. Matthias Tschoep, Heinrich Bassler Registergericht: Amtsgericht Muenchen HRB 6466 USt-IdNr: DE 129521671 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1