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western blot bands wider at the middle than the top


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2 replies to this topic

#1 swatcats

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Posted 24 May 2011 - 02:30 PM

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

#2 Inmost sun

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Posted 25 May 2011 - 04:03 AM

Mw and Rf do not behave linear but exponential in non-gradient gels, so your observation reflects this


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



#3 mdfenko

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Posted 26 May 2011 - 12:24 PM

your samples are meeting more resistance in the 10% gel than in the 7.5% gel and so are spreading more as they travel.
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