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JoVE Encyclopedia of Experiments
Encyclopedia of Experiments: Biological Techniques

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The Luciferase Assay: A High-Throughput Assay to Measure Luminescent Signals Using Engineered Nanoluciferase System-Based Bioreporters

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To evaluate the luminescent signal from the bioreporter using a luciferase assay, first, prepare the reaction components, and use the supernatant as the source of the bioreporter.

Dilute the LgBiT and substrate to 1x before use. Transfer 50 microliters of the supernatant from each well or tube, to a 96-well plate, and add 50 microliters of 1x LgBiT to each well. Incubate for 5 minutes at room temperature.

Open the luminometer software. Add 50 microliters of 1x substrate to each well. Place the plate in the luminometer and run the software.

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