SuperSignal West Femto Maximum Sensitivity Substrate

September 29, 2017

University of South Alabama
Cancer Biology
Graduate Student

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Company:

ThermoFisher Scientific

Product Name:

SuperSignal West Femto Maximum Sensitivity Substrate

Catalog Number:

34095

This enhanced chemiluminescent (ECL) substrate is used for the detection of proteins from Western blotting using a secondary antibody with horseradish peroxidase (HRP). The substrate can detect low femtogram levels of protein. It comes as a kit with two solutions: the peroxide solution and the luminol/enhancer solution.

Experimental Design and Results Summary

Application

Western Blotting

Starting Material

MIA PaCa-2 pancreatic tumor cell lysates

Protocol Overview

After incubation your membrane with a secondary antibody that is linked to horseradish peroxidase (HRP) and washing the membrane adequately, prepare your femto solution. Mix equal parts of the peroxide solution with the luminol/enhancer solution, and add the mixture on top of your membrane that has been blotted dry of any wash buffer. Incubate the membrane in the solution for 5 minutes, blot the membrane dry, and then image the solution using your imager of choice.

Tips

Be sure to incubate the membrane in the solution at least 5 minutes for the maximum effect.

Results Summary

This reagent allows for detection of very low levels of protein by western blotting that can then be imaged using your imager of choice.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Extremely sensitive, economical, stable for long periods of time, easy to use.

The Bad

It is so sensitive that the signal can be extremely intense when your protein of interest is present in high amounts.

The Bottom Line

This femto detection kit is very effective for detecting protein levels in western blotting, especially of your protein of interest is present in low amounts. The reagents are very stable and are very effective.

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