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" <c...@accelagen.com>
To: CCP4BB@JISCMAIL.AC.UK
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 <CCP4BB@JISCMAIL.AC.UK> On Behalf Of Nicola Evans
Sent: Wednesday, February 13, 2019 10:46 AM
To: CCP4BB@JISCMAIL.AC.UK
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?)

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