Must Have Instrument for Imaging Blots, Gels, Cells, etc.

Grambling State University
Biological Sciences
Assistant Professor

Overall

Performance

Ease-of-Optimization

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Company:

Li-Cor

Product Name:

Odyssey CLx

Catalog Number:

9140-01

We use western blots and in-cell westerns to analyze changes in protein expression or modification in tissues and cells. We have almost completely transitioned from ECL to near-infrared detection because it works so well.

Experimental Design and Results Summary

Application

Western blot detection

Starting Material

Rat liver tissue and rat hepatoma cell lines

Protocol Overview

The Odyssey is used with near-infrared secondary antibodies for western blot detection and so-called in-cell westerns in which cells are permeabilized to take up the antibodies. It's as simple as using a scanner. Lay the blot, gel, plate, or whathaveyou on the glass panel, set the detector intensity, and press scan.

Tips

Watch out for non-specific bands if multiplexing antibodies and use the LiCor strip buffer to strip and reprobe.

Results Summary

This is a great system. No more fiddling with ECL reagents and camera exposure times. The infrared signal remains stable for days so you can image when convenient. Sensitivity and linearity equals or betters ECL. Interestingly, some antibodies that didn't work well with ECL are fine with this system. Multiplexing red and green dyes on one blot has been useful, particularly for comparing phosphorylated vs total. The image shows an anti-ubiquitin primary antibody plus a red IR dye-conjugated secondary antibody.

Features Summary

Besides imaging blots, gels (protein/DNA), and cells, this can image small animal and tissue

Additional Notes

None

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Summary

The Good

Versatile imager with stable, sensitive, multiplex signal

The Bad

Instrument is expensive.

The Bottom Line

This is a must-have instrument if you're running a lot of blots.

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