If Comparing Film and Digital Imagers, Expose Blot on Digital Imager First.

If you are trying to compare how the same chemiluminescent Western blot looks when imaged on a digital imager (like the C-DiGit® Blot Scanner) with how it will look when imaged on film, it’s important to know that you should expose the blot to film BEFORE imaging on a digital imager.

Why does this matter? Digital imaging requires capturing the most photons being generated, which is typically immediately after a 5-minute chemiluminescent substrate incubation. Time may be more of an issue with some substrates. For more information on how film and digial imaging compare, read Western Blot Analysis: Comparison of film and the C-DiGit Blot Scanner.

In Table 1 below, performance differences of a Western blot detected with SuperSignal® West Pico when the same blot is imaged over time. Blot was incubated 5 min in substrate before imaging on the C-DiGit Blot Scanner. Images are normalized to the LUT of the optimal blot.

Table 1 Optimal Blot Unsatisfactory Blot Unsatisfactory Blot
Images Optimal Blot with SuperSignal West Pico Unsatisfactory Blot with West Pico Unsatisfactory Blot with West Femto
Conditions: Immediately after incubation with SuperSignal West Pico 26 min after incubation 51 min after incubation
Imaging Time Immediately after incubation with SuperSignal® West Pico 26 min after incubation 51 min after incubation
Scan Setting High High High
Performance LOD – 625 ng, Signal – 338 LOD – 625 ng, Signal – 114 LOD – 625 ng, Signal – 32.2

In Table 2, Performance differences of a Western Blot detected with SuperSignal West Dura1 when the same blot is imaged over time. Blot was incubated 5 min in substrate before imaging on the C-DiGit Blot Scanner. Images are normalized to the LUT of the optimal blot.

Table 2 Optimal Blot Satisfactory Blot Satisfactory Blot
Images Optimal with West Dura Satisfactory with West Dura Satisfactory with West Dura
Conditions:
Imaging Time Immediately after incubation with SuperSignal West Dura 24 min after incubation 48 min after incubation
Scan Setting High High High
Performance LOD – 156 ng, Signal – 12,300 LOD – 156 ng, Signal – 10,400 LOD – 156 ng, Signal – 9,090

In Table 3, Performance differences of a Western Blot detected with SuperSignal West Femto when the same blot is imaged over time. Blot was incubated 5 min in substrate before imaging on the C-DiGit Blot Scanner. Images are linked to the LUT of the optimal blot.

Table 3 Optimal Blot Satisfactory Blot Satisfactory Blot
Images Optimal Blot with West Femto Satisfactory Blot with West Femto Satisfactory Blot with West Femto
Conditions:
Imaging Time Immediately after incubation with SuperSignal West Femto 24 min after incubation 48 min after incubation
Scan Setting High High High
Performance LOD – 156 ng, Signal – 11,500 LOD – 156 ng, Signal – 8,120 LOD – 156 ng, Signal – 6,860

1Comparable to WesternSure® PREMIUM Chemiluminescent Substrate

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Coming in 2013! LI-COR® C-DiGit® System – Digital Film for Chemiluminescent Western Blot Imaging

Are you tired of waiting in line for the darkroom? Or spending your precious budget monies on all that film? OR having to do multiple exposures to get just the right image for publication? Then, you need DIGITAL FILM!

But what, you may be asking, is digital film? Well, that’s the LI-COR C-DiGit System! Image your chemiluminescent Western blot and get great images for publication the FIRST time!