How to elute Ab from Protein G-Agarose ?

Scott Craig s.craig at student.unsw.edu.au
Tue Oct 13 05:48:12 EST 1998


Dear Sidney,
I use a buffer of 0.5M HAc adjusted to pH3.4 with NH4Ac to elute protein G
from an IgG sepharose column, so this should work in the reverse process of
purifiying IgG. Pierce also sell an elution buffer that works at higher pH
but I dont remember what is in the buffer (I think it was high molar MgCl?)
Hope this helps.

Scott

Sidney Cambridge wrote:

> Hi, everybody,
>
> I need to "purify" a purchased antibody from the soup of other proteins
> it came in and thought ProteinG coupled to agarose would be a good idea.
> However, I heard only of one protocol that uses 200mM glycine ph 2.8.
> While I don't care about the pH, I can't have any primary amines in the
> elution buffer for the subsequent experiments. I understand that one can
> dialyse the purified stuff but I'd rather not given that I have little
> Ab to begin with. So, are there any elution buffers that do not contain
> glycine or other primary amines ?
>
> Thanks a bunch.
>
> Sidney






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