kits for coupling protein A sepharose to IgG

Richard J. Shaw shawr at
Fri Feb 17 09:09:16 EST 1995

In article <95039.153616LVM3 at> Lakshmi Madabusi <LVM3 at> writes:
>Date: Wed, 8 Feb 1995 15:36:15 EST
>From: Lakshmi Madabusi <LVM3 at>
>Subject: kits for coupling protein A sepharose to IgG

>I am interested in covalently coupling protein A sepharose to antibodies.  Ther
>e are a lot of kits which allow this, but I would like to get a feedback on the
> experience others have had with these kits. Could you please enlighten me on y
>our experience with any of the   kits used for this purpose? Thanks for your re

We have used a Pierce kit but substituted Pharmacia Sepharose Protein G, as 
our mAbs don't bind to Protein A.  The system worked OK.  See the paper 
Schneider et al, 1982, The J. of Biological Chemistry, v257, n 18, pp 10766, 
as many of these kits are based on this paper.  See also Harlow and Lane, 
Antibodies; A Lab Manual.
Make sure that the antigen preparation you are going to use contains no 
antibodies that bind Protein A (or G) as these will bind to unfilled sites.  
These columns leak only minimal amounts of antibody.

Richard Shaw                          Internet: ShawR at
Researcher (Science technician)
P.O.Box 40063, Upper Hutt 

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