antibody crossreactivity problem
(by ucgatan from ucl.ac.uk)
Tue Jul 3 05:57:25 EST 2007
On Mon, 2 Jul 2007, Pow Joshi wrote:
> On 7/2/07, Michael Sullivan <mlsulliv from wisc.edu> wrote:
> > I've prepared a rabbit polyclonal antiserum against a plant protein of
> > interest. From immunoblots, it appears that the antiserum is
> > crossreacting with the large subunit of Rubisco to make a big fat
> > band.
> Another way would be to preadsorb the serum/ antibody with a bacterial
> "powder".... Molecular cloning methods by Sambrook et al or methodss in
> molecular biology or even the manual "Antibodies" would give you a
> protocol to prepare the bacterial "powder". You don't have to express
> the Rubisco there just yet, I imagine.
I'm not sure i understand; you preadsorb the antibodies against plain _E.
coli_ lysate? How does this remove anti-rubisco specificities?
Tom Anderson, MRC Laboratory for Molecular Cell Biology, UCL, London WC1E 6BT
(t) +44 (20) 76797264 (f) +44 (20) 76797805 (e) thomas.anderson from ucl.ac.uk
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