6x his tag protein purification

Dept of Microbiology bfeilmei at iastate.edu
Fri Mar 26 09:29:12 EST 1999


Help!!

I have cloned GFP behind a 6X his tag to purify the protein.  I am binding
it to TALON Cobolt beads.  The protein almost binds too well, in that, I am
having a lot of trouble eluting it off.  I have heard rumor of someone using
histidine in the elution buffer to compete for the removal of the his tagged
protein.  Does anyone know of a good way to elute hard to remove his tagged
proteins?

I appreciate the help!!





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