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