Lucifersae Assay

November 03, 2015

Texas Tech University Health Science Center
Internal Medicine
Postdoctoral Researcher

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Company:

Promega

Product Name:

Dual luciferase reporter assay system

Catalog Number:

E1910

Reporter systems are generally used to study eukaryotic gene expression. These dual reporters are generally used for experimental accuracy. Basically here we can express as well as measure the two individual reporter enzymes within a single system. It is used for a variety of applications like reporter assays, transcription factors, mRNA processing and protein folding.

Experimental Design and Results Summary

Application

Promoter activity by luciferase assay

Starting Material

Mouse/Human cell lysate

Protocol Overview

Prepare the lysate by using passive 1Xlysis buffer. Transfer into tube or vial. Add 100 microliters of LAR II into the luminometer tube. Program the luminometer and then add the 20 microliters of PLB lysate mix into it. Measure the luciferase activity. Dispense the Stop&Glo reagent and measure the Renilla luciferase activity.

Tips

After adding 1X passive lysis buffer, subsequent freezing and thawing is necessary.

Results Summary

This reporter is used to reduce the experimental variation between the activity of the experimental reporter and the activity of the internal control caused by transfection efficiency.

Additional Notes

None

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Summary

The Good

It is very easy to perform and the results are very accurate.

The Bad

It is very expensive.

The Bottom Line

This dual reporter assay system is used for more reliable interpretation of the experimental data by reducing extraneous influences. This assay also provides rapid quantitation of both reporters either in transfected cells or in cell-free transcription/translation reactions.

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