Hello,
I am facing an issue with my polyacrylamide gel for western blot. This is 12% gel and I have used precast gel and made it with the recipe provided by BIO-RAD. I have been using the same method, recipe and voltage for last 2 years. It used to run smoothly but all of the sudden this problem appeared. Can you suggest what could be the issue?
Thanks in advance!
Oof, that is an unfortunate-looking gel. I'd say there is a problem with the electrodes in the gel rig.
My guess is that the polymerization is starting too early and non-homogenously, making some clumps even before pouring the gel into the casts. Probably the APS has higher concentration than what you think. If I were in your shoes, I would slow-down the polymerization by using maybe half the amount APS ( you might have to go even lower). Alternatively, you can use less TEMED and make sure that it’s added just before pouring the gel into the cast.
it happened to me twice. And it turned out to be the issue with running buffer.
Try a fresh running buffer and keep an eye on the gel while its running that it does not get short of running buffer. As sometimes the buffer on top is a bit foamy and not liquidy.