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antibody purification

Dave Parcej parcej at biophys.NOSPAM.mpg.de
Tue Jan 25 10:58:48 EST 2000

hold on! you said in your first post that the antibodies were unlikely to
be derived from an impurity in the sample. Still, I spose you could blot
and cut out the band to use as the affinity ligand. :)

In article <388DB6B3.B07C6DFB at uni-konstanz.de>,
Frank.Fackelmayer at uni-konstanz.de wrote:

> Dave Parcej wrote:
> > Alternatively, affinity purify your antibody. the easiest way is to adsorb
> > your protein onto PVDF and after blocking (eg with dried milk), incubate
> > with the serum. You can easily elute with 0.2M glycine. HCl pH2.5 but
> > remember to neutralise with 1M tris immediately!! This works well and
> > normally reduces the overall background compared to crude serum
> > Dave
> >
> Unfortunately that does not work very well in removing anti-E.coli
> antibodies, because the protein preparation used for purifying the antibody
> is usually also from recombinant origin and therefore contains bacterial
> proteins. Of course, making affinity purified antibodies is always a good
> idea for improving results. In some cases, it may however be helpful to
> include a immobilized E.coli lysate column as an additional purification
> step.
> Frank

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