I am optimising a new protocol for SDS-PAGE using 40% acrylamide 29:1 solution from Bio-Rad across different percentages. I casted two 6% equivalent gels according to the protocol I have optimised, then started the electrophoresis using the gels. I placed the gels in two electrodes then run them for 75V in 2hr. After the electrophoresis has finished, I stained the gels using Instant Blue stain from Abcam for 30 minutes then imaged them.
I found the visible line formed across the gel as pointed in the image. This issue has been there ever since I started my optimisation experiments. I suspected it is the excess contamination from the running buffer so I changed the buffer in the chamber every time, but the problem persists.
Can anybody help? Thank you
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