Maximum binding of proteinA-fusion protein to IgG agarose

Wolfgang Schechinger wgschech at med.uni-tuebingen.de
Sat Oct 18 10:27:27 EST 1997


Dear Aikiwa, 
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 
whatever appropriate

4) Elute AB with Glycine HCl pH2 (should be aprox 100 mM if I 
remember right

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?

Good luck!

Wolfgang




> 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
-----                              
Wolfgang Schechinger         
University of Tuebingen
email: wgschech at med.uni-tuebingen.de
http://www.medizin.uni-tuebingen.de/~wgschech/research.htm
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