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Western blot bands look like dumbells - (Aug/25/2005 )

Just wondering if anyone had any insight into a problem I'm having... Lately, when I've been running my gels, the bands always appear as dumbells (with strong signal on the edges and often nothing in the middle). I tried seeing whether it was the running of the gel itself, the transfer, or ECL (pierce supersignal). When I stain an untransferred gel by Coomassie, the dumbells still appear, so I'm convinced its either a protein problem or something of this nature. I generally run 5ug or so of protein and use 10% gels.

Anyone have any insight?


You know, I have had the dumbell effect myself. Have no idea what causes it.

Make new protein solubilization buffer.. see if that helps.


Run the gel at a lower voltage and or add more loading buffer. I usually electrophorese at 70V for 90-120 min