Why do the markers drift after transferred to the PVDF membrane?

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Ziyan Su

By the way, the electrophoresis is in 80V and 120V with SDS-page 4%-20% gel, the transfer is in 300mA for 1h. The PVDF membrane is 0.22um.

John Hardy Lockhart

It looks like something is wrong with your transfer setup. All the marker dyes are being pulled to the same point in the top middle of your gel. Something there acts as a stronger cathode than the rest of the cathode during your transfer. Perhaps the insulation on one of the wires has degraded.

Lubna Therachiyil

One reason could be that you are pressing the blot transfer unit so tightly that the gel is smashed inside. How does the protein band look like?

John Hardy Lockhart thank you! I didn't think of this reason before.

Lubna Therachiyil The protein bands are like this picture shows. the gel looked integrated after transfer. And I only add 2 pieces of filter paper in the sandwich. could that be too loose?