HIS TAG - Metal affinity column
junk at hgmp.mrc.ac.uk
Fri Mar 21 07:50:41 EST 2003
Historians believe that in newspost
<da9848bd.0303201841.19f44949 at posting.google.com> on Thu, 20 Mar 2003,
Sandra & Thomas Haselhorst <haselhorst_2000 at yahoo.de> penned the
following literary masterpiece:
> But my
>protein doesn't bind to the matrix at all. I only find my protein in
>the through flow and elution of the metal column with 150 mM Imidazole
>results in several other bands. But not my protein. Is there anything
>wrong with the matrix, vector etc. ?? I am not sure where to start to
>solve this problem ...
It could simply be that the His tag is folded up in such a way in your
protein that it is inaccessible to the column.
Is it N-terminal or C-terminal tagged?
Is your protein a multimer?
What buffer are you using to load the protein on the column?
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
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