Thanks to everyone who responded.
Matthew
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 the original proteolytic site, I have been unable to get cleavage despite using a lot of TEV at 37 C, pH 8.0. Has anyone been able to overcome a similar problem?
Matthew Merski
Post-doctoral researcher
UCSF