Odyssey Clx For Western Blot Imaging

Colorado State University
Food Science and Human Nutrition
Graduate Student

Overall

Performance

Ease-of-Optimization

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Company:

Licor

Product Name:

Odyssey Clx

Catalog Number:

9140-09

The Odyssey Clx is used for western blot imaging. The protocol is very similar to conventional western blotting. It calls specifically for Millipore PVDF membranes, and Odyssey blocking solution used for blocking, primary and secondary incubations. You can use any primary antibody, however, the secondary antibody is specific to Licor, it is conjugated to a fluorophore in order to image. The Odyssey images based on near-IR fluorescence. The Odyssey can image western blot (NIR), in-cell western, gel shift, and tissue sections.

Experimental Design and Results Summary

Application

Western Blotting

Starting Material

Rat hepatoma whole cell lysates

Protocol Overview

Whole cell lysates were separated via SDS-PAGE, and transferred to a PVDF membrane (Millipore). Membrane was blocked in 1:3 OB:TBS for 1 hour at RT. Membrane was probed for b-actin at a 1:1000 dilution in 1:3 OB:TBST (0.1%tween20) overnight at 4 degrees C. Membrane was washed in and incubated with secondary antibody, IRDye680 goat-anti-rabbit 1:15,000. in 1:3 OB:TBST (0.1%tween20) with 0.02% SDS for 1 hour at RT.Membrane was imaged using auto intensity.

Tips

Load very small amount of protein marker, I use Licor chameleon at 1uL.

Results Summary

Nice crisp results. Imaging on this computer are quick and easy. Can image the same blot forever after you dry it out.

Features Summary

Can multiplex antibodies! Saves tons of time

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

EASY, saves time, user friendly

The Bad

Tough to strip membranes

The Bottom Line

Would rather do this any day than chemi!!!

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