Weird Western Blot (circular streaks)

gs_stones at my-deja.com gs_stones at my-deja.com
Thu Jul 20 12:39:48 EST 2000


In article <8l3ns5$45d$1 at nnrp2.deja.com>,
  gs_stones at my-deja.com wrote:
> Hi,
> I've had a couple of weird western blots lately,
> snip

Thanks to folks that have replied already, its good to know that
support is available when things don't work as expected.  With
reference to some of the posts, I've had this problem a couple of times
now, sometimes worse then others, the streaking I refer to appears on
ponceau red stained blots, also on the finished (antibody probed) blot,
and appears to me to be formed of the protein from the gel that has
somehow been swirled? during transfer. As I mentioned before, the gel
runs fine and Coomassie staining the gel after the blot has gone weird
reveals a perfect gel.  The worse occurence of this problem happened
after I used a colleagues blotting apparatus and buffer (I'm told that
the buffer can be used twice) so I used her sponges and blotting rig.
Her blot was fine, mine grossly distorted over about one half of the
blot, so I think that means the apparatus was clean.  This problem has
also happened with fresh blotting buffer.  The only thing I can come up
with is some incompatibility between the gel and the blotting
chemistry, or a problem with non uniform heat distribution over the
blot.  We don't use a stirring bar in the blotting tank, but I'll give
it a go now.
Once again , many thanks for suggestions about this perplexing problem,
if anyone is sufficiently interested I could email a picture of the
blot, no one here has ever seen anything like it!
regards
g s stones


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