Allows Infrared Imaging of HRP-Probed Blots

March 21, 2019

Princeton
Molecular Biology
Graduate Student

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Quality of Results

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Company:

Li-cor

Product Name:

WesternSure substrate

Catalog Number:

926-80020

A western blot was performed normally probing with an HRP-conjugated secondary antibody. This reagent was then applied for 2 minutes to develop. Blot was imaged on a Li-cor Odyssey after, showing good labeling of bands but with some background.

Experimental Design and Results Summary

Application

Western blot

Starting Material

Blocked and probed western blot

Protocol Overview

After washing with TBS-T for 3 times on the final step, this reagent is added in the same way as normal chemiluminescent solutions.

Tips

This reagent permanently labels the membrane in the 680 nm range

Results Summary

Results showed good replication of normal HRP or using an IRDye680 antibody; however, the background was higher with this kit.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Works great to detect low abundance proteins by infrared

The Bad

Higher background than normal chemiluminescent or infrared westerns

The Bottom Line

Works well for very low abundance proteins or bad antibodies. Otherwise, use normal infrared IRDye antibodies instead.

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