how to cleave His-tag

V. Yee vcyee at u.washington.edu
Mon Jul 15 23:29:53 EST 1996


We'd like to engineer a cleavage site in our His-tagged protein
but cannot use a thrombin, fXa, or enterokinase site since these
are calcium-requiring enzymes and we need to keep calcium from 
our protein.  Does anyone have a suggestion for a non-calcium
dependent protease that we can use, and which has been successfully
used by others (we are considering trypsin and chymotrypsin, but
are likely to have cleavage at more than the one desired site)?

Many thanks!





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