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). This is a flash-type assay that requires luminescence to be measured immediately after adding luciferase detection reagents to your sample.
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.
Note: for assaying purified recombinant firefly luciferase in the Passive Lysis Buffer included in the kit, we recommend adding Firefly Luciferase Stabilizer (catalog no. 30083) to the lysis buffer. We also offer the Firefly & Renilla Luciferase Single Tube Assay Kit for Purified Luciferase (catalog no. 30084), which includes all the components of the regular Single Tube Assay Kit plus Stabilizer. Stabilizer is not required for use with cell lysates or purified recombinant Renilla luciferase.
The stated number of assays is based on 96-well plate format. The kits include enough 5X Passive Lysis Buffer to perform the stated number of assays in plate sizes ranging from 96-well to 24-well. If needed, additional 5X Passive Lysis Buffer (catalog no. 99912, see below) can be purchased separately.
- 5X Passive Lysis Buffer
- Firefly Luciferase Assay Buffer 2.0
- Renilla Luciferase Assay Buffer 2.0
- Aquaphile Coelenterazine
- High sensitivity and linearity
- Convenient single tube format
- Water soluble substrates
More Luciferase Kits
|30085||Firefly Luciferase Assay Kit 2.0||Flash-type firefly luciferase assay kit.|
|30075||Firefly Luciferase Assay Kit (Lyophilized)||Firefly luciferase assay kit with lyophilized assay buffer for economical room temperature shipping and convenient storage at -20°C.|
|99923||5X Firefly Lysis Buffer||Extra lysis buffer for use with Firefly Luciferase Assay Kits, only required if using well sizes larger than 24-well.|
|30028||Steady-Luc Firefly HTS Assay Kit||Glow-type firefly luciferase assay with signal half-life of about 3 hours for high-throughput screening in multi-well plates.|
|30028L||Steady-Luc Firefly HTS Assay Kit (Lyophilized)||Steady-Luc kit with lyophilized assay buffer for economical room temperature shipping and convenient storage at -20°C.|
|30082||Renilla Luciferase Assay Kit 2.0||Flash-type Renilla luciferase assay kit.|
|30081||Firefly and Renilla Luciferase Single Tube Assay Kit||Assay kit for measuring firefly and Renilla activities sequentially in the same sample in a single tube.|
|99912||5X Passive Lysis Buffer||Extra lysis buffer for use with Renilla and Dual Luciferase Kits, only required if using well sizes larger than 24-well.|
|30020||ATP-Glo Bioluminometric Cell Viability Assay Kit||Flash-type luminescence-based assay for measuring cell viability or ATP concentration.|
|22003||Mini Cell Scrapers||Mini-cell scrapers, useful for lysing cells in 96-well, 48-well, or 24-well plates.|