Luciferase Assay
Luciferase Assay - Related Videos
02:01
Assessing Pseudovirus Infection Using Luciferase Reporter Assay
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2026
Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021).This video demonstrates a luciferase-based assay to quantify pseudovirus infection in epithelial cells. The pseudovirus encodes the luciferase gene, which is expressed in infected cells. After washing, and cells are incubated with lysis buffer at an ultra-low temperature, and then thawed to induce lysis.
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.
03:44
Luciferase Assay to Study Efficacy of Antiparasitic Chemicals Against Parasites
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2025
The video demonstrates the luciferase assay used to determine the efficacy of an antiparasitic chemical inhibitor against the intracellular pathogen Toxoplasma gondii. The study involves luciferase-expressing parasites invading the host cells to replicate and survive. The chemical inhibitor binds with the cysteine proteases of the parasite and inhibits replication. The luciferase activity detected is proportional to the fold changes in parasite growth, determining the drug's efficacy against...
02:36
The Luciferase Assay: A High-Throughput Assay to Measure Luminescent Signals Using Engineered Nanoluciferase System-Based Bioreporters
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2025
The video demonstrates the luciferase assay used to quantify luciferase-tagged proteins. The assay uses an engineered luciferase enzyme composed of a large fragment that binds with the small fragment, which is conjugated with a protein of interest. The bound subunits form an active enzyme that, in the presence of the substrate, releases a luminescent signal. This signal is then quantified using a luminometer.
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...
FAQs
The luciferase enzyme reacts with its specific substrate, producing photons as a reaction output. A luminometer detects the resulting luminescence, and the measured signal provides a quantitative readout rather than a visual observation. Because signal intensity generally reflects the amount of enzyme or reporter activity present, changes in light output can indicate differences between biological conditions.
A luciferase reporter gene can connect promoter behavior to a measurable luminescent signal. When researchers use this arrangement in cells, changes in promoter activity can be assessed through differences in light production. This creates a quantitative way to examine regulatory mechanisms and determine how experimental conditions affect transcriptional control without relying only on indirect biological observations.
Changes in luminescence can reflect differences in the amount of luciferase enzyme or in reporter activity. The interpretation therefore depends on what biological feature the reporter represents, such as promoter activity, gene expression, or signaling pathway activity. Comparing signal intensity across experimental conditions helps identify whether those processes increase or decrease under the tested circumstances.
A typical measurement requires a luciferase-based reporter system, its specific substrate, and a luminometer. Researchers examine the reporter activity in cells, allow the enzyme-substrate reaction to generate photons, and then measure the emitted luminescence. The recorded signal can be used to quantify biological activity and compare results across the experimental conditions being investigated.
Researchers use this approach when they need a sensitive, quantitative readout of promoter activity or gene expression. A reporter gene places these regulatory events in a form that can be measured through luminescence. The resulting data support comparisons among experimental conditions and can help clarify how biological regulatory mechanisms influence cellular activity.
Researchers can compare luminescence from treated and untreated cellular conditions to assess changes in reporter activity. Depending on the reporter design, the signal may reveal effects on gene expression, promoter activity, or signaling pathways. This makes the assay useful for evaluating cellular responses to drugs or other treatments through a sensitive quantitative measurement of biological activity.