Just to let you know. No way, Talon also don't work. I am gonna try the GE His-trap column.

Fred wrote:
> Hi everyone,
> Thanks for answer my question. Just to add some more notes regarding to my expression system. The insert-vector (pET28) has been sequenced and the his-tag is N-terminal. The anti his-tag WB is positive and the binding buffer's pH is 8.2 (double-checked). > I had already experienced the same problem before, which I solved just increasing urea from 6M to 8M. Now, I have reached GndHCl 6M and no binding at all. > I'm currently running a SDS-PAGE with samples eluted from Talon and let you know the results.
> All the Best,
> Fred
>>
>>
>> --- Fred /<ccp4bb.l...@gmail.com>/ schrieb am *Di, 27.1.2009:
>> *
>>
>>     *Von: Fred <ccp4bb.l...@gmail.com>
>>     Betreff: [ccp4bb] [OFF TOPIC] his-tag doesn't bind
>>     An: CCP4BB@JISCMAIL.AC.UK
>>     Datum: Dienstag, 27. Januar 2009, 22:00
>>
>>     *
>>
>>     *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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