On this episode of Molecular Moments, Chad Briscoe speaks with Dr. David Berkowitz, Director of the Division of Chemistry at
The LI-COR Odyssey Western blot delivers quantitative accuracy for target protein detection by using the platform’s infrared technology to match or exceed chemiluminescent detection sensitivity. Leveraging this platform’s capabilities, BioAgilytix is able to perform accurate, reproducible, quantitative Western blot assays.
LI-COR is an infrared (IR) laser-based instrumentation that is able to detect Western blots without the need for film, a darkroom, or any chemiluminescent substrates. Using infrared fluorescent signals that are directly proportional to the amount of target protein, LI-COR Odyssey can accurately and reproducibly measure changes in protein levels via ratiometric detection. In fact, the platform is able to detect both faint and strong bands within a single image, without signal saturation.
The LI-COR imager yields very low background fluorescence, which enables detection of low-picogram targets with NIR fluorescence, as well as low abundance proteins at endogenous levels. It also enables the detection of two targets simultaneously on the same membrane, which greatly increases accuracy of quantification and provides for more robust comparison.
The LI-COR platform has broad applicability for quantitative Western blots, protein gel imaging, and in-cell Westerns, providing for quantitative analysis and a wide linear dynamic range that is not available with traditional chemiluminescent methods. The accuracy and linearity of the system’s infrared detection provides for confident analysis of the differences in protein expression, and have made LI-COR the standard for quantitative Western blots compared to film.
In addition to two-color infrared and in-gel quantitative Western blots, BioAgilytix uses our LI-COR Odyssey platform for cell-based assays, protein detection, nucleic acid detection, protein arrays, RNAi analysis, and more.
The core benefits of the LI-COR Odyssey stem from its infrared fluorescence technology, which enables superior sensitivity, quantification, and linear range when compared to chemiluminescence and visible fluorescence detection. Its near-infrared detection generates low background fluorescence for a signal-to-noise ratio that is better than visible fluorescence, and equal to or greater than chemiluminescence. In fact, LI-COR is able to accurately detect both strong and weak signals in a single image over a 4-5 log linear dynamic range.
Another key advantage of the platform is its ability to multiplex two different targets on the same blot. Not only does this save time and precious sample volumes by multiplying data output, it makes normalization against an internal control for quantification more precise. This mitigates the potential errors that would be introduced from protein loss during stripping and re-probing, or by comparing blots separately. With its two-color detection and quantification, the LI-COR Odyssey can be used to analyze post-translational modifications of proteins, confirm protein identity, and more.
How do you decide which platform to go with in support of your project? Afshin Safavi, Ph.D., of BioAgilytix offers his expert opinion in this video.
When quantitative Western blot detection or near-infrared fluorescent imaging is needed, the BioAgilytix team knows the LI-COR platform will deliver the sensitivity and linearity required to yield accurate results. LI-COR removes the need for enzymatic reactions and substrates, with highly stable signals that generate high-quality data.
Let our experts show you how we can apply LI-COR’s robust detection capabilities to your next project.