Simulated monolclonals?

Michael Lassner mikel51 at mother.com
Sat Jun 14 08:38:25 EST 1997


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



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