Maximum binding of proteinA-fusion protein to IgG agarose
wgschech at med.uni-tuebingen.de
Sat Oct 18 10:27:27 EST 1997
I purify IgGs from Serum on ProtA agarose using the usual protocol:
1) I dilute serum 10 times with PBS, TrisBuffer or 0.9% NaCl
(whatever is around)
2) Pass through column
3) wash with plenty (10 vol) of buffer, if necessary, watch A280 or
4) Elute AB with Glycine HCl pH2 (should be aprox 100 mM if I
Should work on chemically immobilized AB, too.
Pharmacia has nice N-hydroxy-succinimide activated prepacked columns
(1 and 5ml). Not shure if they're right for your purpose, but I you
have some handy, why not try them out?
> Dear researchers,
> Does anyone know the condition for maximum binding of
> proteinA-fusion protein to IgG agarose? When I used a spent medium
> of transfected cells as a source for the purification, I failed to
> purify it. Does it need to adjust pH to the optimal condition for
> binding to IgG agarose?
> Welcome any comments and suggestions.
> Jun-ichi Aikawa, Ph.D.
> University of California, San Diego
usual disclaimers apply
University of Tuebingen
email: wgschech at med.uni-tuebingen.de
!Junk mail is *not* appreciated!
SPAMs will be answered automatically by sending my 128MByte virtual memory file.
This is *NO* joke.
More information about the Methods