Dual-glo Luciferase System
Dual-glo Luciferase System - Related Videos
12:55
High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
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Cited by 45 •
2014
We present a rapid and inexpensive screening method for identifying transcriptional regulators using high-throughput robotic transfections and a homemade dual-glow luciferase assay. This protocol rapidly generates direct side-by-side functional data for thousands of genes and is easily modifiable to target any gene of interest.
07:33
In vivo Dual Substrate Bioluminescent Imaging
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Cited by 20 •
2011
Herein we describe the methods to construct, visualize, and quantify the bioluminescent reactions of both firefly and renilla luciferase enzymes expressed in metastatic breast cancer cells during their growth and metastasis in vivo.
13:48
Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay
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Cited by 6 •
2008
Here, we describe procedures for studying changes in phagocytosis-induced gene expression with a luciferase-based reporter gene approach using the Dual-GloTM Luciferase Assay System from Promega.
04:07
A Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-Infected Cells
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2025
This video demonstrates an assay to study the translation regulation in poxvirus-infected cells. Uninfected and virus-infected cells were co-transfected with mRNAs encoding for firefly luciferase (Fluc) and Renilla luciferase (Rluc), with the Fluc mRNA containing a 5' Poly(A) leader sequence. Upon adding bioluminescent substrates on lysed cells, measure the signal from both the luciferases to identify the translational advantage of the Fluc mRNA in virus-infected cells, conferred by the leader...
05:51
Construction of CRISPR Plasmids and Detection of Knockout Efficiency in Mammalian Cells through a Dual Luciferase Reporter System
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2020
Here, we present a protocol describing a streamlined method for the efficient generation of plasmids expressing both the CRISPR enzyme and associated single guide RNA (sgRNAs). Co-transfection of mammalian cells with this sgRNA/CRISPR vector and a dual luciferase reporter vector that examines double-strand break repair allows evaluation of knockout efficiency.
FAQs
Firefly luciferase functions as the experimental reporter, so its activity reflects the gene-regulatory or signaling event under study. Renilla luciferase provides an internal control measured from the same biological sample. Comparing these paired signals helps distinguish changes in the experimental response from variation affecting the sample more generally.
The ratio between the two signals helps compensate for differences in cell number, transfection efficiency, and sample handling. These factors can alter the total light produced without representing a genuine change in gene regulation. Internal normalization therefore makes comparisons between samples more quantitatively reliable than interpreting a single reporter signal alone.
The assay first uses reagents to generate the firefly luciferase light signal. A subsequent reagent selectively quenches that reaction, allowing the Renilla luciferase activity to be measured separately from the same sample. This sequence prevents the two readouts from being treated as one combined signal and supports direct comparison of their activities.
A normalized result reflects firefly reporter activity relative to the Renilla control rather than raw light output alone. An altered ratio can therefore indicate a change associated with the tested regulatory or signaling condition, provided that the control remains suitable for normalization. The paired measurement gives greater confidence when comparing biological samples.
Cells are analyzed for firefly luciferase activity first, using reagents that produce its measurable light signal. The firefly reaction is then selectively quenched, and Renilla luciferase activity is measured from the same biological sample. Researchers compare the two readings, commonly using the control signal to normalize the experimental reporter.
The system is useful when researchers need to evaluate how promoters or enhancers affect reporter activity. Firefly luciferase supplies the experimental readout, while Renilla luciferase helps account for sample-to-sample variation. This paired design supports quantitative comparison of regulatory elements and can help validate their influence on gene expression.
Changes in the normalized reporter signal can be used to examine transcriptional regulation or cellular signaling within a biological experiment. The firefly measurement reports the experimental response, and the Renilla measurement provides context for interpreting that response. As a result, the system can help analyze pathway-related effects while reducing the influence of handling and transfection differences.
A single reporter cannot independently reveal whether signal differences arise from the tested regulatory process or from variation in cell number, transfection efficiency, or sample handling. The Dual-Glo Luciferase System adds an internal control from the same sample, making normalized comparisons more reliable for promoter, enhancer, pathway, and regulatory-element studies.