Initiation any GST-fusion protein and His-tag protein purification?
AEVDOKIMOZ at cinci.rr.com
Mon Nov 25 09:13:30 EST 2002
> very unstable after cleavage with thrombin, can be only stable in PMSF in
So don't use such a promiscuous protease. Clone it with TEV or other 3C-type
protease site, under an MBP.
> The binding efficiency to gluthione sepahrose resin seemly depends on the
> protein expression and folding, which may be affected by some unknown
> such as sonication?
Try French press or microfluidizer instead, if you think that there's a
problem with sonication (there often is!).
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