Why do Western Blot bands appear dumbbell shapes instead of straight lines?

1. The biggest possibility of dumbbells is that the gel is not well configured, and the gel is uneven after solidification. It is recommended to appropriately extend the gel solidification time to ensure the quality of the gel. 

2. It may be that the sample contains too many impurities, and did not centrifuge down, then the impurities are deposited in the middle of the well, and the protein is naturally pushed to both sides. The sample can be centrifuged before loading to allow impurities to precipitate. 

3. Pay attention to whether the film is very hot after the end of the transfer, if the membrane is hot, it needs to be properly cooled.  

Hope these can help you!

1. The most likely reason for the dumbbell bands is that the gel is not properly configured, and the uneven solidification of the gel is likely to appear dumbbell.

 2. Another possibility is that there are too many impurities in the sample, which are not centrifuged, and then the impurities are deposited in the middle of the hole, and the protein is naturally pushed to the sides.