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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   

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Chavas Leonard, Ph.D. @ home
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