Separating GST from GST-fusion protein

Dwayne Taliaferro via methods%40net.bio.net (by taliaferrod from mail.nih.gov)
Mon Apr 20 11:25:37 EST 2009


This is also a problem with MBP protein tags as well.  I had luck using a
ion exchange columns and eluting using salt.

> Getting rid of trace GST after cleavage is not trivial IMO.   After
> expressing a GST fusion protein we cleaved away the GST using a thrombin
> cleavage site and ran an affinity column to get rid of uncleaved fusion
> protein and free GST.  The resulting flowthrough has a single band on
> Coomassie blue stained gel but still has GST present as measured by ELISA.
> We've switched to plan B - a different construct.
> 
> Allison
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