Weird Western Blot (circular streaks)

Bernard Murray, PhD spam at 127.0.0.1
Wed Jul 19 18:05:12 EST 2000


In article <8l3ns5$45d$1 at nnrp2.deja.com>, gs_stones at my-deja.com wrote:

> I've had a couple of weird western blots lately,
> The problem shows up when ponceau red stained as
> weird circular streaks over some (not all) of the
> blot.

Poor transfer (bubbles in the sandwich)?  Protein-contaminated
sponges or cassettes?  Membrane not in a homogeneous state
(uniformly wet with the same buffer) before transfer or
staining?  Dirty staining box?

> Coomassie staining the gel reveals that
> the gel has run ok.  Generally a part of the blot
> is ok, normal bands, but over the rest the lanes
> go into a circular streak which extends over
> several lanes.

I've seen that with; a) grossly overloaded samples, b) samples
with high salt, c) samples containing detergent.  However,
these are distortions within the gel rather than between the
gel and the blot so if your Coomassie stained gel looks good
it is unlikely to be this.
   I hope you don't soak your gel before transfer.  It's a
waste of time and can lead to artefacts.

   Bernard

-- 
Bernard P. Murray, PhD
bpmurray at cgl . ucsf . edu
Department of Cellular & Molecular Pharmacology, UCSF






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