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 normalising transfection control for firefly luciferase assays.
The Firefly and 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. 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.
The Firefly and Renilla Luciferase Single Tube Assay features Biotium’s water-soluble Aquaphile™ coelenterazine. It also includes 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 stabilise the enzymes.