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Description

Firefly luciferase is widely used as a reporter for studying gene regulation and function, and for pharmaceutical screening. It is a very sensitive genetic reporter due to the absence of endogenous luciferase activity in mammalian cells or tissues. Firefly luciferase is a 62,000 Dalton protein, which is active as a monomer and does not require subsequent processing for its activity. The enzyme catalyzes ATP-dependent D-luciferin oxidation to oxyluciferin, producing light emission centered at 560 nm. Light emitted from the reaction is directly proportional to the number of luciferase enzyme molecules. The Firefly Luciferase Assay is designed for simple and efficient quantitation of firefly luciferase reporter enzyme activity from cultured cells with high sensitivity and linearity. This is a flash-type luminescence assay that requires signal to be measured immediately after adding working solution to samples. The luminescence signal decays over the course of about 10 minutes of reaction time, although signal half-life may vary depending on luciferase expression levels.