antibody purification

Frank O. Fackelmayer Frank.Fackelmayer at uni-konstanz.de
Mon Jan 24 09:49:17 EST 2000


Marianne Schwartz wrote:

> I have raised antibody against a protein, which was purified on a Ni2+
> column, after expressed in E.coli. (has His TAGs in the N-terminal). The
> protein looked perfectly pure on a gel, however, the antibody recognize
> some E.coli proteins also, as judge from a Western. . Can I get rid of
> these antibodies ?
> Does anybody has a protocol for this problem.
> Thanks
> Marianne
>
> ---

Hi Marianne,
polyclonal antisera often contain anti-E.coli antibodies. This is usually
not a problem of protein purity used for immunization, but seems to be due
to natural anti-E.coli immune response (in fact, most non-immunized rabbits
already have these antibodies). As this problem is quite common, there is
already a commercial remedy, the immobilized E.coli lysate sold by Pierce
(no affiliation!). It comes with columns to easily purify your serum by
just passing it over the immobilized lysate.

Frank








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