His-tagged proteins....

[Debashis Mukhopadhyay]

Hi All!

This question is directed to those 'gurus' who have
expertise with His-tagged proteins to help me out of
this problem. I'm new to this "affinity tagged"
protein field.

I have a protein domain which is cloned and expressed
in E.coli having an N-terminal His-tag and a Thrombin
cleavage site just after that. The idea is to get rid
of the tag for crystallisation purposes by thrombin
treatment. Can you point me to any reference(s), ideal
for a novice, having details for the same and
subsequent purification steps to eliminate the
contaminants! Any nice protocol would as well be good!

Any help would be highly appreciated.
Regards,
Debashis

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