I am working with the bacterial his tag expression system from Qiagen, and am trying to purify the his-tag protein. Apparently, the recombinant protein is going into inclusion bodies, so I am trying to purify it under denaturing conditions (8 M urea). The problem is that after lysis there is a lack of the His tag, because it is not longer recognised by Ni-NTA conjugate (Qiagen) Does any one have any recommendations of how to generate a lysate without losing the His tag?
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