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Luciferase Reporter Assay

A molecular biology technique used to study gene expression and regulation. It involves the use of luciferase, an enzyme that produces bioluminescence, as a reporter. This enzyme is encoded by a reporter gene that is placed under the control of a regulatory sequence of interest. When the regulatory sequence is activated, luciferase is expressed, and its activity can be measured through a luminescent signal. This assay is widely used for investigating promoter activity, studying cellular signaling pathways, and in drug discovery.

Workflows requiring the Luciferase Reporter Assay

Gene Promoter Activity Analysis

In the study of gene regulation, the Luciferase Reporter Assay is pivotal for analyzing promoter activity. Researchers use this assay to link luciferase expression to specific promoter sequences, enabling them to quantify how different factors, such as transcription factors or external stimuli, influence gene expression. This approach is fundamental in understanding gene regulation mechanisms and in identifying potential therapeutic targets.


Signal Transduction Pathway Studies

By constructing reporter genes that respond to specific signaling pathways, scientists can monitor the activation and inhibition of these pathways in real-time. This method is particularly valuable in dissecting complex cellular signaling mechanisms and in drug discovery, where it’s used to screen for compounds that modulate these pathways.


High-Throughput Screening for Drug Discovery

The Luciferase Reporter Assay allows researchers to rapidly assess the effects of thousands of compounds on specific gene expression pathways. This application is crucial for identifying potential drug candidates that can modulate disease-relevant genes, streamlining the early stages of drug development.


Methods of Performing a Luciferase Reporter Assay

Single Luciferase Assay

The Single Luciferase Assay is a simpler method, focusing on the activity of one luciferase enzyme, typically the firefly luciferase.



Dual-Luciferase Reporter Assay

The Dual-Luciferase Reporter Assay is a sophisticated method that measures the activities of two different luciferases (commonly firefly and Renilla) in a single sample. This approach allows for internal normalization, providing more accurate and reliable data by correcting for variations in cell viability and transfection efficiency.



Live-Cell Luciferase Reporter Assay

The Live-Cell Luciferase Reporter Assay is designed for real-time monitoring of luciferase activity within living cells. This method is particularly useful for dynamic studies of gene expression and cellular responses over time.


Luciferase Reporter Assay workflows have never been easier

The OT-2 is a bench-top liquid handler designed to be accessible and flexible enough to automate many common applications.

How to Automate the Luciferase Reporter Assay process:


Benefits of Automation over Manual Pipetting for Luciferase Reporter Assay:

Protocol Spotlights

Opentrons helps you automate Luciferase Reporter Assay with open-source protocols for the OT-2 and Opentrons Flex


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