Smiling bands (curved bands) (Western Blot)

Question

Accepted Answer

Bands curve upward at the edges of the gel, appearing like a smile — the middle of the lane runs faster than the sides. Common causes: 1) Voltage too high during electrophoresis 2) Running time too long 3) Gel or buffer overheated during the run 4) Poor heat dissipation in the gel chamber Solutions: 1) Reduce voltage to 80-120V; longer runs at lower voltage give better resolution 2) Shorten run time — stop when the dye front reaches 2/3 of the gel 3) Pre-chill running buffer; run with an ice pack against the chamber, or pause to cool 4) Use a larger buffer volume, or perform the run at 4 C in a cold room