I had a similar pb with Mark 12 from Invitrogen. I was told to boil my standard before loading and it did help a lot. Cba
From: proteins-bounces at oat.bio.indiana.edu on behalf of Steve G
Sent: Fri 9/22/2006 4:04 PM
To: proteins at magpie.bio.indiana.edu
Subject: [Protein-analysis] Larger Proteins Vanish During Gel Run
This problem was recently related to me by several protein-running
members of my lab. I don't know where to begin looking for answers for
such an intermittent problem in both pre-cast and self-made SDS-PAGE
gels, so I thought I'd ask here.
As the gel runs, the visible Bio-Rad dual color protein standard bands
are seen to start to separate, later the larger ones (100-250 kD) start
to fade, then vanish. The smaller ones seem fine, dye front fine too.
Post-transfer Ponceau S stain shows there was a loss of protein in the
sample lanes in that same upper size range as well - just like the
marker. Smaller proteins and smaller markers are all there.
Since this happens to the commercial marker (in it's own loading
buffer) and the protein/lysate samples (in our own), in both precast
and self-cast gels, the only thing in common would seem to be the
electrophoresis buffer - but that's made in large volumes and used for
multiple gels - and the problem doesn't show all the time...
Thanks for any comments.
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