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Introduction to Quantitative Infrared Westerns
Figure 1. Time course of JNK phosphorylation, in response to LPS treatment. Phospho-JNK is shown in green, total JNK in red. Yellow indicates overlapping signals. Reprinted with permission from Bond, D. et al. Biol Proced Online 10(1): 20-28 (2008).
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Use infrared fluorescence, IRDye® secondary antibodies, and Odyssey® imagers to detect Western blots without film, a darkroom, or any chemiluminescent substrates.
- Signal is directly proportional to the amount of target protein
- Quantify both low and high-abundance proteins across the widest possible linear dynamic range
- Matches or exceeds chemiluminescent detection
- Exceeds visible fluorescence detection
- No enzymatic reactions, substrates, or timing concerns
- Fluorescent signals are stable indefinitely — image when YOU are ready
- Save time by not doing multiple exposures to get "just the right image"
- Detect two different targets on the same blot
- Easy normalization against an internal control, for increased accuracy
- No stripping and reprobing of blots
- Monitor changes in protein expression or stability over time — even subtle changes
- Measure protein phosphorylation and cell signaling events
- Validate RNAi or proteomic studies
- Perform quantification of protein abundance