Atp-based Luciferase Assay
Atp-based Luciferase Assay - Related Videos
08:32
The ATP Bioluminescence Assay
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2023
In fireflies, the luciferase enzyme converts a compound called luciferin into oxyluciferin, and produces light or “luminescence” as a result. This reaction requires energy derived from ATP in order to proceed, so researchers have exploited the luciferase-luciferin interaction to gauge ATP levels in cells. Given ATP’s role as the cell’s currency of energy, the ATP bioluminescence assay can provide insight into cellular metabolism and overall cell health.In this video, JoVE discusses cellular...
03:24
ATP-Based Luciferase Viability Assay: A Homogenous Method to Evaluate the Growth-Inhibitory Potential of Test Agents on Tumor Organoids
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2025
In this video, we demonstrate a rapid, single-addition assay to assess cell viability in patient-derived organoids upon treatment with a test agent using intracellular ATP levels as a marker for cellular metabolic activity. The intracellular ATP levels are measured using the luciferase/luciferin reaction, where the emitted light intensity indicates the cellular viability and, in turn, the test agent's growth-inhibitory potency.
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.
03:34
Luciferase-Based Growth Assay to Quantify Intracellular Parasite Reproduction
0 Views •
2025
In this video, we demonstrate the luciferase assay to quantify the growth of the intracellular pathogen Toxoplasma gondii. Utilizing a strain of Toxoplasma that expresses luciferase enzyme after infecting the host cells, the luciferase activity is detected in the infected cell culture to determine the successful reproduction of the parasite.
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.
FAQs
Each component supports the light-producing reaction: luciferase catalyzes the reaction, luciferin serves as the light-emitting substrate, and oxygen and magnesium ions provide required reaction conditions. ATP participates in this enzyme-catalyzed process, so changing its concentration changes the number of photons produced. Maintaining these components under controlled conditions helps make luminescence comparisons meaningful.
Living cells maintain ATP pools, whereas damaged or nonviable cells contain substantially less ATP. Consequently, the measured light signal can serve as an indirect indicator of the amount of biologically active cellular material in a sample. Higher or lower luminescence supports quantitative comparisons of viability, proliferation, or metabolic activity when the assay conditions are consistent.
Signal reliability depends on controlling the reaction environment, particularly the availability of luciferin, oxygen, magnesium ions, and ATP, because each contributes to photon production. Differences in these conditions can alter luminescence independently of the biological variable being studied. Consistent treatment of samples therefore supports valid comparisons between experimental groups and more dependable interpretation of ATP differences.
A basic workflow begins by preparing the biological samples, combining them with the luciferin and luciferase reaction components, and maintaining the required oxygen and magnesium conditions. The resulting luminescence is then detected and compared across samples. Using the same handling and measurement conditions throughout the experiment helps relate signal differences to ATP content rather than procedural variation.
The assay provides a rapid light-based readout and is compatible with microplate formats, allowing many samples to be measured and compared within a screening experiment. This format is useful when researchers need quantitative comparisons across conditions, such as differences in cell proliferation or metabolic activity. Its sensitivity further supports detection of ATP-associated changes among tested samples.
Because living cells maintain ATP, the assay can detect ATP-associated biological material in samples where microbial contamination is a concern. A luminescence measurement provides a rapid quantitative signal that can be compared between samples or conditions. In biological techniques, this makes the method useful for screening contamination alongside its roles in studying cell viability and proliferation.
Differences in luminescence can support quantitative comparisons of ATP levels among samples, which may reflect differences in cell viability, proliferation, or metabolic activity. The signal does not independently identify which biological process caused the change, so interpretation should remain connected to the experimental context. This makes the assay valuable for comparison-based studies and screening rather than interpretation in isolation.