Imager Sensitivity Settings May Affect Detection of Chemiluminescent Western Blot Signals

Standard and High Sensitivity Settings on the C-DiGit
Standard and High Sensitivity Settings on the C-DiGit
Making sure that the sensitivity setting is optimal to capture the most signal from your chemiluminescent Western blot could be the difference between getting a good, strong signal or getting a signal that you can barely see. This is our possible cause 7 for weak chemiluminescent signals.

How can you avoid possible cause 7 for LI-COR chemiluminescent imagers? On the C-DiGit® Blot Scanner, use High Sensitivity setting (12-min scan) for more sensitive detection. On the Odyssey® Fc Dual-Mode Imaging System, use a longer integration time (up to 10 min). Why is this important? Well, digital imaging with the C-DiGit Blot Scanner or Odyssey Fc Imager will not generally reach a saturation point. Begin with a longer acquisition time to ensure best sensitivity, then optimize to shorter scan times.

In Table 1 below, we tested the performance differences of a Western blot detected with SuperSignal® West Dura on the C-DiGit Blot Scanner when the same blot is imaged on High Sensitivity (12 min scan) versus Standard Sensitivity (6 min scan). As you can see, the longer scan time and higher sensitivity make a big difference in the results.

Table 1 Optimal Blot Satisfactory Blot
Images Optimal Sensitivity Setting on C-DiGit Satisfactory Chemiluminescent Western Blot
Conditions: SuperSignal West Dura1 SuperSignal West Dura
Sensitivity High (12 min) Standard (6 min)
Performance Signal – 12,300 Signal – 5,030

1Comparable to WesternSure® PREMIUM Chemiluminescent Substrate

So be sure to check your sensitivity settings before you scan!

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