Firefly & Renilla Luciferase Single Tube Assay Kit

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Product Description

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.


Kit components:

  • 5X Passive Lysis Buffer
  • Firefly Luciferase Assay Buffer 2.0
  • D-Luciferin
  • Renilla Luciferase Assay Buffer 2.0
  • Aquaphile Coelenterazine

Features:

  • High sensitivity and linearity
  • Convenient single tube format
  • Water soluble substrates

Assay schematic

 

 

Example of Firefly & Renilla Luciferase Single Tube Assay using lysates from untransfected HeLa cells (No tfx), or cells transfected with either firefly luciferase alone or co-transfected with firefly and Renilla luciferases. Firefly signal is the luminescence from cells transfected with firefly luciferase alone after adding firefly working solution, while Quenched Firefly is the residual luminescence after adding Renilla working solution to the same reaction. Renilla signal is the luminescence measured after adding Renilla working solution to a firefly reaction with cells co-transfected with firefly and Renilla luciferases.

Example of data from the Firefly & Renilla Luciferase Single Tube Assay using lysates from untransfected HeLa cells (No tfx), or cells transfected with either firefly luciferase alone or co-transfected with firefly and Renilla luciferases. Firefly signal is the luminescence from cells transfected with firefly luciferase alone after adding firefly working solution, while Quenched Firefly is the residual luminescence after adding Renilla working solution to the same reaction. Renilla signal is the luminescence measured after adding Renilla working solution to a firefly reaction with cells co-transfected with firefly and Renilla luciferases.

 

More Luciferase Kits

Catalog numberProductDescription
30085Firefly Luciferase Assay Kit 2.0Flash-type firefly luciferase assay kit.
30075Firefly Luciferase Assay Kit (Lyophilized)Firefly luciferase assay kit with lyophilized assay buffer for economical room temperature shipping and convenient storage at -20°C.
999235X Firefly Lysis BufferExtra lysis buffer for use with Firefly Luciferase Assay Kits, only required if using well sizes larger than 24-well.
30028Steady-Luc™ Firefly HTS Assay KitGlow-type firefly luciferase assay with signal half-life of about 3 hours for high-throughput screening in multi-well plates.
30028LSteady-Luc Firefly HTS Assay Kit (Lyophilized)Steady-Luc kit with lyophilized assay buffer for economical room temperature shipping and convenient storage at -20°C.
30082Renilla Luciferase Assay Kit 2.0Flash-type Renilla luciferase assay kit.
30081Firefly and Renilla Luciferase Single Tube Assay KitAssay kit for measuring firefly and Renilla activities sequentially in the same sample in a single tube.
999125X Passive Lysis Buffer Extra lysis buffer for use with Renilla and Dual Luciferase Kits, only required if using well sizes larger than 24-well.
30020ATP-Glo™ Bioluminometric Cell Viability Assay KitFlash-type luminescence-based assay for measuring cell viability or ATP concentration.
22003Mini Cell ScrapersMini-cell scrapers, useful for lysing cells in 96-well, 48-well, or 24-well plates.

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Additional Information

Size

50 assays, 150 assays, 1,000 assays

MSDS (PDF):MSDS 30081

Protocol (PDF): PI-30081

Shipment Method: Shipping and handling methods will be assessed and calculated at time of shipment based upon item(s) storage temperature conditions.

Expedited shipment may be requested at time of checkout.

Please note that products with recommended storage at 4°C or -20°C may ship at ambient temperature. This will not affect product performance. When you receive the product, place it under the recommended storage conditions.

More information about product handling and shelf life