Help, my negative control is positive

Richard Buick Richard.Buick at
Fri Dec 13 19:14:33 EST 2002

Hi Peter
Your polyclonal antibody has been raised against a recombinant protein.
Purification of this recombinant protein is rarely 100% successful. I would
guess that the recombinant prtein is mammalian, and therefore not as
immunogenic as any bacterial proteins that have managed to "hitch a ride" as
far as injection into the bunnies. Your band that is being detected is a
bacterial protein that is present in untransformed cells, the "purified"
recombinant protein, and control cells. It is unfortunate that it may be the
same size as your recombinant protein. I would suggest further boosts of
recombinant protein in the bunnies.



Richard J. Buick

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"Peter" <krijger at> wrote in message
news:d091ebf2.0212131153.723728b9 at
> Hi,
> Every time i do a Western Blot using a polyclonal antiserum against a
> recombinant protein i get a band in my negative lane. It is the same
> band as in the lane with only the recombinant protein and the postive
> cell controll. It is the only band i see. Does someone know the
> answer??
> thanks..

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