Dear Fred --
just to check... are you sure you have the His-tag? Might have been
cleaved somehow? You might want to increase the number of His in the
tag as well.
HTH.
-- Leo --
On 27 Jan 2009, at 21:00, Fred wrote:
Hi ccp4 list,
I am trying to purify a his-tag protein by metal affinity
chromatography. The protein was expressed in inclusion bodies and
its his-tag doesn't bind the Qiagen Ni resin in denatured
conditions (urea 8M or GndHCl 6M). Playing with NaCl and detergents
didn't help much.
Any help is appreciated.
Fred
------------------------------------------------------------
Chavas Leonard, Ph.D. @ home
Research Associate
Marie Curie Actions Fellow
------------------------------------------------------------
Faculty of Life Sciences
The University of Manchester
The Michael Smith Building
Oxford Road
Manchester Lancashire
M13 9PT
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Tel: +44(0)161-275-1586
e-mail: leonard.cha...@manchester.ac.uk
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