help! binding to nickle column

Richard J. Dudley rjdudley at YAHOO.COM
Fri Aug 14 07:45:15 EST 1998


---liz <liz at METHYLGENE.COM> wrote:
>
> I am trying to bind a 180 KDa protein with a His tag to a nickle
column
> (invitrogen) but most of the protein comes out in the flow through.  I
> have tried both native and denaturing conditions.  Can anybody offer
any
> suggestions?

Make sure there is no EDTA nor ionic detergents in your protein
suspension.

Also, search these archives at www.bio.net; this problem comes up a lot.

rich
==
--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley at iname.com)
Research Specialist V
Dept. of Cell Biology and Physiology
The University of Pittsburgh
->biosci archives can be searched at www.bio.net <-

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