STREP-Tag purification

Philipp Wechner philipp.wechner at uibk.ac.at
Mon Jun 24 11:50:30 EST 2002


>
>
> It might be that the His-tag prevents it from proper refolding, but
> perhaps you didn't try hard enough finding a suitable buffer.

I don't think that the his tag prevents propper folding - because i came to the
same results refolding the untaged protein.

>
>
> What have you tried so far? What do you know about the protein - size,
> pI, are there refolding data about homologues?

size is about 20 kDa, pI at 9.1, no data about homologues (i am sure about this!)

i tried different pH for refolding - it worked best with 1,2 M Tris at pH7.3
other buffers (MOPS, HEPES, and about 10 others) were not able to induce refolding

i did a lot of different buffers and i am very happy that i found the buffer i am
using now (and it took me nearly a year). I do not want to go back and do
refolding experiments - i think it is easier to find a tag that makes it able to
purify my protein under these conditions.


>
>
> Bye, Frank
> --
> > But I don't really see running SETI at Home as practical as Folding at Home.  What,
> > exactly, would be the benefit of finding intelligent aliens on the other side
> > of the galaxy?
> Maybe they're broadcasting the principles of protein folding... [from bionet.*]




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