> The DTT is not the biggest problem. I want to extract my protein from
inclusion
> bodies in E.coli. After treating the IB with 7M Urea or 5 M GuHCl i tried
to
> get my protein by NTA-Chormatography (C-terminal 6xHis tag). However, the
> protein does not bind too good - but its OK.
I understand your plight. The best you can do is try - there is no telling
if STREP tag will work in your case, unless you dialyze into better buffer.
have you considered fusing your protein to a thermophilic MBP, and trying to
renature the fusion while keeping MBP alive ? PfuMBP does not really
denature in 5.5 M guanidinium hydrochloride at normal temperature, your
protein probably does...
A.G.E.
