'Sticky' His-tag proteins

Yoram Gerchman yoramg2 at LEONARDO.LS.HUJI.AC.IL
Fri Jul 24 04:44:25 EST 1998


Hi

>Has anyone experienced a 'sticking' of His-tag proteins to
>agarose/Sepharose/any sort of beads at all, causing problems with
>purification (even NTA-agaroes beads that have been stripped of Ni++
with
>EDTA). Ideally I want to keep things 'native'. Phararmacia suggested
using
>a borate buffer to reduce non-specific interactions but this didn't
seem to
>help.

>Any suggestions would be greatly appreciated

>Rob Layfield
>University of Nottingham

I'm not sure if it will help but we purify our protein in the present of
500mM NaCl. It might "shade off" some charges in protein/agaros.




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