western blot bands wider at the middle than the top - (May/24/2011 )
Hi
Maybe this is very basic...but I can't figure this right now. When I use a 7.5% precast gel, the marker bands run quite smooth but when I do the same thing on the 10% gel the marker bands are smoother at the top but seem to get wider at the bottom. I have now noticed this for a couple of times only with the 10% gels. Both are from BioRad and are brand new. Could anything be there with the buffer not mixing well? Or the voltage? I use the tris-glycine-SDS buffer (10x diluted to 1x as with the other gels) and the voltage is always between 90-110.
Thanks
Mw and Rf do not behave linear but exponential in non-gradient gels, so your observation reflects this
swatcats on Tue May 24 22:30:18 2011 said:
Hi
Maybe this is very basic...but I can't figure this right now. When I use a 7.5% precast gel, the marker bands run quite smooth but when I do the same thing on the 10% gel the marker bands are smoother at the top but seem to get wider at the bottom. I have now noticed this for a couple of times only with the 10% gels. Both are from BioRad and are brand new. Could anything be there with the buffer not mixing well? Or the voltage? I use the tris-glycine-SDS buffer (10x diluted to 1x as with the other gels) and the voltage is always between 90-110.
Thanks
your samples are meeting more resistance in the 10% gel than in the 7.5% gel and so are spreading more as they travel.