Immuno-precipitation

Hiromi Tagoh htagoh at hotmail.com
Sat Jan 15 07:54:44 EST 2000


"Beate Fisslthaler" <Fisslthaler at em.uni-frankfurt.de> wrote in message
news:01BF5EAF.3A70EC30 at zphys7205.klinik.uni-frankfurt.de...
> Is there anybody out there...
> who can help me??
> Is there a possibility after a immuno-prcipitation to dissociate the
anti-gen from the anti-body without the dissociation of the two chains of
the anti-body. My antigen I have to detect is exactly 55 kD in size and I
have by every method I used the antibody on my western-blots.
> Thanks for your help and ideas
> Beate.
> ---

I don't understand your aim properly, but I think it depends on what you
want to do after the immuno precipitation.
Anyway,
In affinity purification using monoclonal/polyclonal antibodies, we use
acidic condition to dissociate molecule from insolubilized antibodies.  The
antibodies are re-usable, so the molecule should be intact after the
dissociation treatment.  But usually we use plenty amount (several ml) of
dissociation buffer (not 50 microL ), so if you want to detect the molecule
by PAGE or something, you might have to concentrate the solution.
Any help?





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