protein electrophoresis

Paul Klosen klosen at neurochem.u-strasbg.fr
Wed Apr 26 09:33:58 EST 2000

Edward wrote:
> Hi, this might be a silly question, but I really need your help.
> I am using the product of immunoprecipitation for electrophoresis, that is,
> the samples are antibody-antigen complexes (is that right?).
> My question is that will there be a possiblity of degrading, or breaking of
> the ab-ag complexes, during the procedure of SDS-PAGE, which is a denaturing
> electrophoresis.
> Thanks in advance.
> Ed

Denaturing SDS-PAGE will most certainly dissociate ab-ag complexes. If
your sample buffer includes beta-mercaptoethanol or dithiotreitol, it
will even separate your antibodies into light and heavy chains. SDS-PAGE
after immunoprecipitation is not normally used to analyse the complexes
themselves, but rather to identify the subunits of these complexes.

Paul Klosen, PhD
CNRS UMR 7518 Neurobiologie des Fonctions Rythmiques et Saisonnieres
Universite Louis Pasteur  12, rue de l'Universite F-67000 Strasbourg,
Tel.  Fax.  klosen at neurochem.u-strasbg.fr

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