We have the same experience with the DO NOT AGITATE. We purify our own His-tagged TEV protease, and flash-freeze it IMMEDIATELY after IMAC prep at about 1 mg/mL.
Laurie Betts UNC On Thu, Mar 31, 2011 at 10:37 AM, Charles Allerston < charles.allers...@sgc.ox.ac.uk> wrote: > I echo this. > > Some time ago I was working on a target which seemed to precipitate when > cleaving overnight with TEV. I wasted a fair bit of time trying to optimise > cleavage conditions with a myriad of buffers. In the end, just by not > agitating my solution, there was no precipitation and recovery was extremely > good. > > I never agitate my solutions when cleaving now and have not had any > problems with cleavage (to do with this issue, anyway) or recovery. > > > cheers > charlie > > > > Dr. Charles Allerston > Genome Integrity Group > Structural Genomics Consortium > Nuffield Department of Medicine > Old Road Campus > University of Oxford > OX3 7DQ > http://www.sgc.ox.ac.uk/ > > > > > > -----Original Message----- > From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of > Xiaopeng Hu > Sent: 31 March 2011 15:23 > To: CCP4BB@JISCMAIL.AC.UK > Subject: Re: [ccp4bb] Question about TEV cleavage > > Our experience is do not shake the tube during TEV cleavage,I dont know > why, but it does help. > > xiaopeng >
