10 Possible Causes of Weak Signals on Chemiluminescent Western Blot Images

Are you seeing weaker than expected (hoped for. . .) signal on your chemiluminescent Western blot images with your digital imager? 

Not sure what could be causing this? 

Well, here is a list of 10 possible reasons why you might be seeing weak signals in chemiluminescent Western blot data:

1. The chemiluminescent substrate does not have a fast enough rate of reaction.
2. Not enough substrate was added to the blot.
3. Membrane was placed on the detection system incorrectly.
4. Blot was not detected or processed on the same day it was imaged.
5. Blot was not kept uniformly wet during the entire image acquisition.
6. Blot was exposed to film BEFORE imaging on a digital imager.
7. Blot was imaged using incorrect sensitivity setting.
8. Chemiluminescent substrate was too cold.
9. Chemiluminescent substrate was not incubated for 5 minutes.
10. Substrate was diluted.

Hum, that is quite a list! 

For details on ways to eliminate or avoid these causes and get great results with your chemiluminescent Western blots, read Good Westerns Gone Bad: Maximizing Sensitivity on Chemiluminescent Western Blots.

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