Firefly luciferase has been used as a sensitive reporter to study a wide range of biological responses. However the change of the expression of Firefly luciferase reporter can be due to a global effect instead of a specific effect. The accuracy of Firefly luciferase reporter can be improved by normalizing to a control reporter, such as Renilla luciferase reporter, in the same sample.
The Dual Luciferase (Firefly-Renilla) Assay System is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture. The Firefly Luciferase Reagent is first added to the cells in medium directly. This reagent lyses the cells and contains a substrate for firefly luciferase to produce firefly luciferase luminescence. Next, the Renilla Luciferase Reagent is added to the same well. It quenches the firefly luciferase luminescence and provides the substrate for renilla luciferase to produce renilla luciferase luminescence. The light production of both reactions can be conveniently measured on a luminometer. This assay system has several features:
• Sensitive – highly sensitive detection of firefly luciferase activity and Renilla luciferase activity.
• Stable – the luciferase signal output is stable for more than one hour, providing flexibility with regard to incubation time
• High-throughput – simple homogeneous protocol minimizes handling steps to support high-throughput screening applications
• Compatibility – works well with a variety of common media containing 0-10% serum and phenol red.