How to elute Ab from Protein G-Agarose ?

John Thompson jrt at home.com
Tue Oct 13 17:20:11 EST 1998


I modified the elution buffer Scott described (0.5M HAc adjusted to
pH3.4 with NH4Ac), same pH but only 50 mM HAc. Works fine for eluting
protein A from IgG sepharose.  Haven't tried it with protein G.  My
guess is you could use NaOH to adjust the pH if you want to stay away
from amines.
Regards,
John Thompson

sidney at neuro.mpg.de (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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