I once cleaved a GST tag on the resin using TEV by rocking overnight at 4 C. I would say it is 100% cutting judging from the gel. One thing to add is that the protein bound so tight to the beads that cutting tag is the only way to elute it except by SDS. I haven't had any trouble with TEV and thrombin with rocking or shaking. The important part is that you have to have good protease to start with.
Nian Huang, Ph.D. UT Southwestern Medical Center On Thu, Mar 31, 2011 at 2:41 PM, gauri misra <kamga...@gmail.com> wrote: > Just an offshoot of the same Question.. > I would like to ask whether the same applies for GST-tag digestion using > thrombin.. > No agitation gives better results in the above case too... > Any personal experiences.... > > On Thu, Mar 31, 2011 at 11:29 AM, Klaus Piontek < > klaus.pion...@ocbc.uni-freiburg.de> wrote: > >> And not at full moon! >> >> Klaus >> >> >> Am 31.03.2011 um 16:23 schrieb Xiaopeng Hu: >> >> Our experience is do not shake the tube during TEV cleavage,I dont know >> why, but it does help. >> >> xiaopeng >> >> >> Dr. Klaus Piontek >> Albert-Ludwigs-University Freiburg >> Institute of Organic Chemistry and Biochemistry, Room 401 H Albertstrasse >> 21 >> D-79104 Freiburg Germany >> Phone: ++49-761-203-6036 >> Fax: ++49-761-203-8714 >> Email: klaus.pion...@ocbc.uni-freiburg.de >> Web: http://www.chemie.uni-freiburg.de/orgbio/w3platt/ >> >> >
