Luciferase Substrate Detection
Luciferase Substrate Detection - 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.
11:44
Surface Enhanced Raman Spectroscopy Detection of Biomolecules Using EBL Fabricated Nanostructured Substrates
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Cited by 26 •
2015
We describe the fabrication and characterization of nano-biological systems interfacing nanostructured substrates with immobilized proteins and aptamers. The relevant experimental steps involving lithographic fabrication of nanostructured substrates, bio-functionalization, and surface-enhanced Raman spectroscopy (SERS) characterization, are reported. SERS detection of surface-immobilized proteins, and probing of protein-ligand and aptamer-ligand binding is demonstrated.
03:38
Measuring Mycobacterium smegmatis Mistranslation Rates Using a GFP/Luciferase Reporter System
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2025
Source: Chen, Y., et al., Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems. J. Vis. Exp. (2019)This video demonstrates a method to quantify mistranslation in Mycobacterium smegmatis using an inducible GFP and secreted mutant luciferase reporter system. It shows how luminescence resulting from mistranslation is measured and compared to GFP fluorescence to evaluate the impact of a test molecule on translational accuracy.
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.
03:49
A Luciferase-Fluorescent Reporter Influenza Virus to Track Viral Infections
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2025
This video demonstrates a method to track viral infections in mouse models using a recombinant luciferase and fluorescence protein-expressing bi-reporter Influenza A virus. This method enables both in vivo and ex vivo studies, enabling researchers to gain insights into viral dynamics and the effects of interventions.
FAQs
The substrate is not merely a reagent; it determines whether the luciferase reaction can generate a measurable signal. Under defined reaction conditions, luciferase catalyzes substrate oxidation, and the resulting photon output is collected as a quantitative readout. Changes in signal can therefore indicate altered enzyme abundance or altered enzymatic activity, depending on the assay design.
Signal intensity must be interpreted in relation to both the reporter and the reaction environment. More light may reflect more luciferase, greater enzyme activity, or changes in conditions that affect the reaction. Keeping reaction conditions defined helps distinguish biological changes from assay-related variation, which is especially important when comparing infection states or immune responses.
Luciferase substrate detection can support either live-cell monitoring or endpoint measurement, and the choice changes the information gained. Live-cell measurements help follow biological activity over time, whereas endpoint assays provide a defined readout at a selected stage. In infection studies, this distinction can separate dynamic host–pathogen behavior from final experimental comparisons.
A typical workflow begins with a biological system carrying or expressing a luciferase reporter, followed by addition of the appropriate substrate under defined conditions. The reaction is then measured with a luminometer, which quantifies emitted photons. These controlled conditions produce a measurable signal that can be compared across samples to assess differences in biological activity.
Within immunology and infection research, the readout can address several biological questions rather than one type of experiment. Reporter activity can indicate gene expression, while luciferase-linked measurements can also track pathogen burden or host–pathogen interactions. This flexibility allows the same measurement principle to examine pathogen behavior and how host cells respond.
Immune-cell responses and therapeutic interventions are two important application areas. A luciferase signal can monitor activity associated with an immune response, while changes during treatment can help evaluate an intervention’s effect on infection-related biology. The method is particularly useful when high sensitivity and a broad dynamic range are needed.