If you would biotinate your IgG before the immunoreaction, you should be
able to pull out the biotinated IgG even under conditions that dissociate
the antibodies from the antigens (e.g.: 0.1N NaOH). For example, in my
BIAcore experiments, the biotinated oligonucleotides stay bound to the
immobilized streptavidin even in 0.1M NaOH. The loss for each cycle is <2%
of the remaining immobilized streptavidin. The stability for the
strepavidin-oligo complex appears higher because we have reycled the
immobilized oligo through up to 97 cycles of 0.1M NaOH and still able to do
protein-DNA binding studies. I have not done low pH conditions. Let me know
if it works also at pH 2.
Hope this helps!
Tony
At 11:48 AM 9/9/98 -0400, Kenneth Williams wrote:
> Is there a reasonably generic means of removing (or at least decreasing)
>immunoglobulins from immunoprecipitates (after release of the bound Ag from
>polyclonal Ab) prior to SDS PAGE purification of the Ag?
>
>
>
>Ken Williams, Ph.D.
>Director, HHMI Biopolymer/Keck Laboratory
>Professor (Adjunct) Research
>Molecular Biophysics and Biochemistry
>
>
>Visit the HHMI Biopolymer/Keck Laboratory Web Page at:
>
> http://info.med.yale.edu/wmkeck/
>
>
>
************************************
Dr. Anthony T. Yeung, Ph.D.
Director, Fannie E. Rippel Biotechnology Facility
Member, Institute for Cancer Research
Fox Chase Cancer Center
7701 Burholme Ave. Philadelphia, PA 19111
Voice: 215-728-2488
FAX: 215-728-3647
email: AT_Yeung@FCCC.edu
http://www.fccc.edu/research/labs/yeung/
************************************