Hi Matthew,
By now, you have received many posts telling you both how efficient and
inefficient TEVp is. You might be confused. This seeming contradiction
can be explained by a few events, among many others: Inaccessibility of
cleavage site, absence of reducing agents, and presence of deterge
Ok, to sum up for the board, a good reference for this problem is at:
http://mcl1.ncifcrf.gov/waugh_tech/faq/tev.pdf
Thanks to everyone who responded.
Matthew
On Mon, May 24, 2010 at 9:27 AM, Matthew Merski <
mer...@blur.compbio.ucsf.edu> wrote:
> Hello all,
>
> I am working with a protein
Hi Matthew,
TEV protease is very robust. I normally digest with 1:100 ratio
according to the OD280. I normally digest at 4C for overnight around 16-18
hours. Make sure your tev protease site are not inaccessible and buried
inside.
best
Xiaohu
On Mon, May 24, 2010 at 12:27 PM, Matthew Merski <
_
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
Matthew Merski
Sent: Monday, May 24, 2010 9:28 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] TEV cleavage problems
Hello all,
I am working with a protein that is expressed as with an N-terminal domain
that is normally
Hello all,
I am working with a protein that is expressed as with an N-terminal domain
that is normally cleaved for activation of the protein (and
crystallization). For in vitro reasons I've needed to switch the normal site
to a TEV site. However, even though the TEV site is in the same place as t
