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:
- The chemiluminescent substrate does not have a fast enough rate of reaction.
- Not enough substrate was added to the blot.
- Membrane was placed on the detection system incorrectly.
- Blot was not detected or processed on the same day it was imaged.
- Blot was not kept uniformly wet during the entire image acquisition.
- Blot was exposed to film BEFORE imaging on a digital imager.
- Blot was imaged using incorrect sensitivity setting (learn about the easy-to-use Image Studio™ Software. Try FREE Image Studio Lite Western Blot Analysis Software to see just how easy it is!)
- Chemiluminescent substrate was too cold.
- Chemiluminescent substrate was not incubated for 5 minutes.
- Substrate was diluted.
Hum, that’s 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.