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.

One step and Two step systems

The TWO-Step 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.

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.

Some data with TWO-Step Luciferase (Firefly and Renilla) assay system

Learn more in these posts on our blog