Rene, There are many proteases that are suitable for digestion of an N-terminal tag. Interestingly, there are NOT many (any?) proteases suitable for digestion of thr C-terminal tag (except for the dual-enzyme His-tag digest system that sometimes work and sometimes does not).
I personally like TVMV, TEV, and Thrombin. With the first two enzymes, you have no worries whatsoever about non-specific digestion, whereas Thrombin is pretty gentle but has been reported to eat things up sometimes. Cheers, Artem > Hi, > > A non-ccp4 Q. Sorry. > > I would like to use a cleavable purification tag at the N-terminus/ > extracellular end of my membrane protein for purification. Before I > start, I wonder if someone could recommend a particular protease site > that I can engineer between the tag and my protein? How about a > proprietary cleavage system such as the PreScission protease (GE > Healthcare)? I would be grateful to hear success and horror stories > in this area. > > Best wishes, > > Rene > > ================================================ > Dr R.A.W. Frank, PhD > Royal Commission for the Exhibition of 1851 Research Fellow > > Prof Seth Grant Lab / Genes to Cognition > Wellcome Trust Sanger Institute > Hinxton > Cambridge CB10 1SA > > Work Tel: 0044 (0)1223 834244 ext. 7318 > Cell No.: 0044 (0)7870 208280 > =============================================== > > > >
