Initiation any GST-fusion protein and His-tag protein purification?

Artem Evdokimov 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
factors,
> such as sonication?

Try French press or microfluidizer instead, if you think that there's a
problem with sonication (there often is!).

Good luck,

A.G.E.





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