antibody purification

Dave Parcej parcej at biophys.NOSPAM.mpg.de
Tue Jan 25 05:38:09 EST 2000


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


In article <388CC36A.815C7397 at post6.tele.dk>, bio-a079 at post6.tele.dk
(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
> 
> ---




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