Could I possibly inquire as to which method you used for blotting the membrane? In my experience, the wet method using tanks has occasionally resulted in a partial missing or non-specific point signal in the detection when SDS is included in the transcription buffer.
The blot on the left is a poor transfer looks like a huge air pocket... between the gel and membrane, the blot on the right could be a pinched or fold in the gel or membrane while maintaining contact with each other. Another possibility is that your gel was compromised in some way.
Yasuhiro Nishida Thanks for the answer. I used the Towbin buffer is 25 mM Tris, 192 mM glycine, pH 8.3 with 20% methanol. It does not contain SDS. I used 100v for 90 min for transferring.
Wenping Wang. Thank you for your comment. I consider this condition to be very common. As Jeffrey P Cheng pointed out, this is most likely due to the formation of air bubbles. The adhesion between the gel and the membrane should be tightened by rolling a disposable thick pipette or similar and using a sufficient amount of filter paper and sponge to ensure that there is no gaps in the cartridge. It is also important to note that the buffer heats up when energized, so it should be cooled sufficiently and agitated so that bubbles do not accumulate in one place. Low pressure may prevent overheating of the buffer.
Good luck.
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