Smeary 2-D protein gels - help!!

Aine L. Plant aplant at sfu.ca
Wed Mar 19 13:52:44 EST 1997


Hello all:

We have been running 2-D gels for approx. 2 years using the protean 2-D
gel system.  Up until now the gels have been running well, however,
recently we have started to periodically obtain gels that have
substantial amounts of smearing.  This happens about every 20 gel runs
and appears to be a problem with the SDS-PAGE rather than the ief since 
1)  it only applies to a few tubes out of the total run for ief and 2)
the markers are also smeared.  We have checked our solutions and are
careful when casting gels.  

These are gels of in vivo labelled proteins.  The second dimensions are
run out with 2 gels per protean apparatus (i.e. we don't double up the
gels) and the run is cooled.

If anyone has any insight or advice we would be very grateful indeed.

Many thanks in advance

Aine L. Plant
Biological Sciences
Simon Fraser University
Burnaby  B.C. V5A 1S6
CANADA



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