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Beschreibung

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. However, the light production resulting from the reaction leads to formation of suicidal adenyl-oxyluciferin at the enzyme surface. It results in very short half-life of the light emission with a flash-type kinetics. The Firefly Luciferase HTS Assay is designed for is a proprietary mixture of substances that modify the enzymatic reaction to produce a long-lasting signal (steady-glow) by preventing the formation of adenyl-oxyluciferin at the enzyme surface. It is a homogeneous high sensitivity firefly luciferase reporter gene assay kit for the quantification of firefly luciferase expression in mammalian cells with signal half-life of about 3 hours (Figure 2). Glow-type luciferase assays have lower luminescence signal compared to flash-type assays. The sensitivity and limit of detection of the assay will depend on luciferase expression levels in your experimental system as well as luminometer sensitivity.