Detection of Firefly Luciferase Activity by Using Luciferin as a Substrate
Principle
Luciferin 4-monooxygenase (ATP-hydrolyzed) extracted from firefly, Photinus Pyralis.
Fluorescein + ATP + O2 → oxygen and worm fluorescein + PPi + H2O + light
The optical density I is related to the Michaelis-Menten equation
where H is the Planck constant and v is the frequency of emitted light. When the ATP concentration is very low ([ATP] < < Km), the Michaelis-Menten equation simplifies to:
I = Constant × [ATP]
Constant = V/Km. optical density is proportional to [ATP].
Appliance
cell-based assay
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Da — when not otherwise indicated, molecular weight units are daltons. Mw — weight-average molecular weight. Mn — number-average molecular weight.
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Cite this article
Aladdin Scientific. "Experiments for the determination of luciferase" Aladdin Knowledge Base, updated Dec 24, 2024. https://www.aladdinsci.com/us_en/faqs/experiments-for-the-determination-of-luc-en.html
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