Flash-type Renilla Luciferase assay

Renilla luciferase has been used as a reporter gene for studying gene regulation and function in vitro and in vivo. It commonly is used in multiplex transcriptional reporter assays or as a normalizing transfection control for firefly luciferase assays. The enzyme does not require post-translational modification for its activity and may function as a genetic reporter immediately following translation. Coelenterazine also emits light from enzyme-independent oxidation, a process known as autoluminescence. The autoluminescence is enhanced by superoxide anion and peroxynitrite in cells and tissues. The Renilla Luciferase Assay utilizes a special buffer formulation designed to yield reliable, linear measurements of Renilla luciferase activity with minimal autoluminescence background and superior sensitivity. 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 2 minutes of reaction time, although signal half-life may vary depending on luciferase expression levels.