Simulated monolclonals?

[Andrei Popov]

Michael Lassner wrote:
> 
> I have raised several antisera to E. coli produced proteins that give
> less than satisfactory results for Western analysis.  There is a
> relatively high level of background bands when using the antibodies
> against E. coli extracts.  I tried affinity purifying the antibodies
> against the his tag purified proteins used to inoculate the bunnies, but
> this didn't make a significant improvement.  My next thought was to have
> some peptide(s) synthesized, link to CnBr sepharose, and affinity purify
> antibodies that react with specific a specific peptide or peptides.  Can
> anybody give me some advice about whether this approach is worth
> trying?  Any suggested protocols?  Any advice about how to pick the
> peptides?  Any suggestions as to a good source of synthetic peptides?
> 
> Thanks in advance,
> Mike








The easiest thing would be to absorb the antisera on a column with
total E.coli proteins (minus your protein of interest).
 
best wishes


-- 
Andrei Popov

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