Firefly and Renilla luciferases are widely used as reporter genes for studying gene regulation and function, and for pharmaceutical screening. Renilla luciferase is often used in conjunction with firefly luciferase as a normalizing transfection control or for multiplex transcriptional reporter assays (1, 2). As with many enzymes, firefly luciferase and Renilla luciferase follow Michaelis-Menten kinetics and thus maximum light output is not achieved until substrates (above the Km) and co-factor are present in large excess. When assayed under these conditions, light emitted from the reaction is directly proportional to the number of luciferase enzyme molecules. Biotium’s Firefly & Renilla Dual Luciferase Assay Kit is designed for detection and quantification of firefly and Renilla luciferase reporter enzymes from cultured cells in a simple, efficient and linear fashion. Note: This is a two-tube dual luciferase assay. Separate aliquots of lysate are required for the firefly and Renilla reactions. For the assay protocols listed below, 20 uL lysate is required for firefly luciferase measurement and 20 uL lysate is required for Renilla luciferase measurement, therefore 40 uL total lysate is required to measure both enzyme activities, equivalent to 2 wells of a 96 well plate. See the product protocol under downloads for more information.
- Sensitivity and Linearity: Linear correlation between reporter gene expression and light output
- Low Autoluminescence & High Sensitivity: Reduced autoluminescence background for Renilla luciferase assay
- Convenience: One kit for both luciferase assays.