SuperSignal West Pico PLUS Chemiluminescent Substrate

October 04, 2017

University of South Alabama
Cancer Biology
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

ThermoFisher Scientific

Product Name:

SuperSignal West Pico PLUS Chemiluminescent Substrate

Catalog Number:

34080

The SuperSignal West Pico PLUS Chemiluminescent Substrate is great for detecting protein levels from western blots that have been incubated with horseradish peroxidase (HRP)-linked secondary antibodies. I used this substrate to detect GST levels in a sample that I probed with a GST mouse primary antibody and an anti-mouse HRP-linked secondary antibody.

Experimental Design and Results Summary

Application

Western Blotting

Starting Material

MIA PaCa-2 pancreatic tumor cell lysates

Protocol Overview

After incubating your membrane with your HRP-linked secondary antibody, wash the membrane thoroughly with your wash buffer of choice. Mix the peroxide and luminol solutions from the Pico kit as a 50:50 mixture. Blot the membrane dry and add enough of the mixture to completely cover the membrane. Incubate for 5 minutes at room temperature, and image the membrane with your imager of choice.

Tips

Be sure to incubate the membrane in the solution at least 5 minutes for the maximum effect, but longer times may be needed if protein levels on your membrane are low.

Results Summary

I was able to detect femtogram levels of my GST fusion protein with the Pico reagent and the BioRad ChemiDoc Imager. There was hardly any background.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Easy to use and detect low levels of protein. Less expensive than other detection reagents.

The Bad

If protein levels are very low, the SuperSignal West Femto Reagent may be better to detect the protein with shorter exposure times.

The Bottom Line

The SuperSignal West Pico PLUS Chemiluminescent Substrate is very effective for detecting proteins from a western blot. It is an affordable alternative to other detection reagents that is sufficient for most blotting applications.

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