Immuno-precipitation

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Fri Jan 14 10:41:56 EST 2000


In article <01BF5EAF.3A70EC30 at zphys7205.klinik.uni-frankfurt.de>,
Fisslthaler at em.uni-frankfurt.de (Beate Fisslthaler) wrote:
> 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.
> ---


Try leaving the reducing agent out of the sample buffer, and don't boil
it. The SDS alone should dissociate the antigen from the Ab, but the
IgG chains should remain associated with each other because of the
disulfide bonds between the chains.

Nick



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