Bands look thick and bloated, edges spill out, strong-signal differences are masked; not suitable for densitometry quantification because the linear range is lost.
Common Causes
1Exposure time too long
2Substrate concentration too high, or target protein expression is very high
3Imaging gain setting too high
Solutions
1Collect short, medium, and long exposures — pick a non-saturated image for analysis
2Reduce loading amount or dilute the antibody appropriately
3Switch to a medium-sensitivity ECL substrate
4Ensure the signal stays within the linear dynamic range of the imaging system when quantifying