This kit is designed to measure Firefly and Renilla luciferase activities in transfected cell reporter assays. 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. 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 Firefly & Renilla Luciferase Single Tube Assay is a combined luciferase assay allowing sequential measurement of Firefly and Renilla luciferase activity in the same sample with high sensitivity and linearity (Figure 1). Firefly luciferase activity is measured first, then Renilla Luciferase Assay Buffer 2.0 is added to the sample to simultaneously quench firefly luciferase activity and measure Renilla luciferase activity. The Renilla Luciferase Assay Buffer 2.0 quenches the firefly luciferase activity to the level of untransfected cells, allowing sequential measurement of firefly and Renilla luciferase activity in the same sample (Figure 2).

Note: This is a flash-type assay that requires luminescence to be measured immediately after adding luciferase detection reagents to your sample. It requires a single-tube luminometer or luminescence microplate reader with reagent injectors.

This kit is a new and improved replacement for the Firefly & Renilla Dual Luciferase Assay Kit (catalog no. 30005). In addition to providing a simplified, single tube assay, the kit features Biotium’s Aquaphile™ coelenterazine, a water soluble substrate that can be stored at -20°C with minimal evaporation, unlike methanol solutions of coelenterazine.

The Firefly & Renilla Luciferase Single Tube Assay features Biotium’s water-soluble Aquaphile™ coelenterazine. It also includes our new 1X Passive Lysis Buffer 2.0, which is ready-to-use without dilution for cell lysis. The buffer also can be used to dilute recombinant firefly or Renilla luciferase enzymes without the need to add BSA to stabilize the enzymes.

The stated number of assays is based on 96-well plate format. The kits include enough 1X Passive Lysis Buffer 2.0 to perform the stated number of assays in plate sizes ranging from 96-well to 24-well. If needed, additional 1X Passive Lysis Buffer 2.0 (catalog no. 99821) can be purchased separately.