Easy to use for protein quantification

University of Nebraska Medical Center
Pharmacology and experimental neuroscience
Graduate student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Scientific Pierce

Product Name:

SuperSignal West Pico Chemiluminescent Substrate

Catalog Number:

34080

This product was used in Western blot assays looking at various proteins in the brain homogenate of MPTP-intoxicated animals. After isolating the region of interest, protein was purified, run on a gel, and transferred to a PVDF membrane. Membrane was developed with equal parts of SuperSignal reagents for 5 minutes and imaged.

Experimental Design and Results Summary

Application

Western blot

Starting Material

Brain homogenate

Protocol Overview

Purify protein before running on a gel and transferring to a PVDF membrane. Mix equal parts of SuperSignal reagents just prior to use (approx 2.5 mls each per membrane) and incubate at RT for 5 minutes.

Tips

May not work well to develop membranes incubated with weak primary antibodies or if the protein of interest is not well expressed.

Results Summary

Used with beta actin primary antibodies, this works great

Additional Notes

None

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Summary

The Good

Easy to use

The Bad

May not work for all antibodies

The Bottom Line

When we need to quantify protein and normalize to beta actin, this is what we use.

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