Western Blot Problem

Daniela Gadotti gadodani at isu.edu
Thu Jan 22 13:10:28 EST 1998



> Help! I am getting a bizarre signal on my western blots.   the signal
> shows up not only as the band I am trying to detect (B-actin) but also as
> vertical streaks in
> between the lanes stretching from the top of the blot to the bottom

Describe the streaks better.  Are they "dotty", "punctuated"?  If so it is
probably a keratin artefact, i.e. :  1:   there is keratin in your reagents
and equipment, and 2:  one or both of your antibodies has a subset of
immunoglobulines that recognize keratin.  If the subset is small, the
problem will only show with very sensitive detection methods, like ECL.
This is common and there are ways to fix it.  Look it up in "Medline"
under western or electrophoresis AND artefacts.

Daniela Gadotti
gadodani at isu.edu

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