Luciferase Glow Reagent
Luciferase Glow Reagent - 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.
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.
10:13
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
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Cited by 6 •
2013
Achieving a systems level understanding of cellular processes is a goal of modern-day cell biology. We describe here strategies for multiplexing luciferase reporters of various cellular function end-points to interrogate gene function using genome-scale RNAi libraries.
12:58
Transfecting RAW264.7 Cells with a Luciferase Reporter Gene
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Cited by 13 •
2015
Transfection into the macrophage cell line, RAW264.7, is difficult due to the cell’s natural response against foreign materials. We described here a gentle yet robust procedure for transfecting luciferase reporter genes into RAW264.7 cells.
10:17
Preparing Anhydrous Reagents and Equipment
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2023
Source: Laboratory of Dr. Dana Lashley - College of William and Mary Demonstrated by: Timothy Beck and Lucas Arney Many reactions in organic chemistry are moisture-sensitive and must be carried out under careful exclusion of water. In these cases the reagents have a high affinity to react with water from the atmosphere and if left exposed the desired reaction will not take place or give poor yields, because the reactants are chemically altered. In order to prevent undesired reactions with H2O...
FAQs
The measured light is the assay’s readout of luciferase reporter activity: greater reporter activity can generate more detectable photons under the same assay conditions. Because the reagent converts enzyme catalysis into a visible or instrument-recorded signal, investigators can compare reporter expression among experimental samples and assess changes caused by infection, host signaling, or treatment.
Luciferin supplies the substrate that luciferase oxidizes, while ATP and oxygen support the reaction in systems that require them. Oxidation produces an excited product, and photon release occurs when that product returns to a lower energy state. These components connect the biochemical reaction to the light signal used for quantitative reporter measurements.
The reporter’s biological placement determines the interpretation. A reporter associated with infected cells or a pathogen-host interaction can indicate infection-related activity, whereas a reporter linked to host signaling can reflect immune responses. Consequently, the same light-producing chemistry can answer different questions, including infection burden, host response, or treatment effects, depending on the assay design.
Visible emission can provide a direct indication that the luciferase reaction is producing light, while an instrument can record the signal for comparisons between samples. Instrument-measured output is especially useful when the goal is to quantify reporter expression or evaluate changes across infected, untreated, or treated experimental conditions.
A basic workflow uses samples containing luciferase reporter activity together with the glow reagent, which supplies the reaction environment for light production. The resulting emission is then observed or measured with an instrument. Comparing signals across appropriately selected samples allows researchers to evaluate reporter expression, infection-related activity, immune signaling, or treatment effects.
In these studies, the reagent supports assays of infected cells, host responses, and pathogen-host interactions. Researchers can use the resulting reporter signal to examine infection burden, monitor immune signaling, or compare treatment effects. This provides a common light-based measurement while preserving flexibility in whether the reporter reflects pathogen-associated activity or a host biological response.