Lane broadening on protein gels

Michael Holland M.Holland at dwe.csiro.au
Sun Nov 21 20:20:41 EST 1993


Hi gurus,
We have problems with unacceptable lane broadening with our samples during
SDS-PAGE. We have to solubilize our material by heating at pH 10 and 70 C
for 10 minutes. The pH is adjusted to 7 with acetic acid and the samples
Speedy Vaced to dryness. The residue is taken up in 20 ul of standard Laemmli
sample buffer at pH 6.8 and loaded onto Hoeffer mini gels. From the colour
of sample buffer, which contains bromophenol blue, the pH remains unchanged.
When the gels runs everything looks great but on staining the bands are 2-3
times the width of the lane. The MW standards run fine as do other gels with
different samples. Clearly the problem seems to be in the sample
preparation. Any hints?????
Michael K Holland PhD
CSIRO, Wildlife & Ecology
PO Box 84, Lyneham ACT 2602, Australia
Fax: 61-6-242-9242  Phone:61-6-242-1793
mholland at dwe.csiro.au



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