Why Do Western Blot Bands Appear Darker at the Edges and Lighter in the Center?

In Western Blot (WB) analysis, bands that are darker at the edges and lighter in the center are often due to sample overloading. When the amount of protein loaded exceeds the gel's capacity, proteins may not migrate evenly during electrophoresis, especially in the central region of the gel. This results in faster or further migration at the edges, making the center of the band appear lighter.

 

To address this issue, reduce the amount of protein loaded onto the gel, ensuring appropriate concentration and volume for more uniform bands. Additionally, verify the electrophoresis and transfer steps to rule out other factors causing uneven protein migration.

 

MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.

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