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Firefly & Renilla Luciferase Single Tube Assay Kit

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.

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30081-1, 30081-2, - 30081-TView allHide

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50 assays, 150 assays, 1,000 assays

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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.

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.


Kit components:

  • 1X Passive Lysis Buffer 2.0
  • 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 or cells transfected with either firefly luciferase alone (Firelfy Only) or co-transfected with firefly and Renilla luciferases (Firefly + Renilla). In cells transfected with firefly only, the Renilla signal is the residual firefly luminescence after adding Renilla working solution to the reaction. Luminescence was measured on a Promega Glomax® 20/20 single tube luminometer using the Dual Glo® program with integration time of 1 second.

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.
999125X Passive Lysis BufferExtra lysis buffer for use with Renilla Luciferase Assay Kits, only required if using well sizes larger than 24-well.
30081Firefly and Renilla Luciferase Single Tube Assay KitAssay kit for measuring firefly and Renilla activities sequentially in the same sample in a single tube.
99821-100mL1X Passive Lysis Buffer 2.0 Extra lysis buffer for use with Firefly & Renilla Single Tube Assay 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.

References

1. Alam, J. and J.L. Cook. 1990. Anal. Biochem. 188:245-254.
2. Bronstein, I., et al. 1994. Anal. Biochem. 219:169-181.
3. Gould, S.J. and S. Subramani. 1988. Anal. Biochem. 175:5-13.
4. Brasier, A.R., et al. 1989. BioTechniques. 7:1116-1122.
5. Bhaumik S. et al. (2004) J Biomed Opt. 9, 578-86.
6. Matijasevic Z. et al. (2001) Carcinogenesis. 22, 661-4.
7. Nieuwenhuijsen BW. et al. (2004) J Biomol Screen. 8, 676-84.
8. Matthews, J.C., Hori, K. and Cormier, M.J. (1977) Biochemistry 16, 85–91.

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Support & faq

Luciferase Assays

No. Biotium’s Renilla and Firefly Assay Kits are based on flash luminescence. Samples should be treated and read individually. Alternatively, you may purchase the Steady-Glo Firefly Assay Kit for HTS. We currently do not have a Steady-Glo for Renilla Assays.

Category: Luciferase Assays

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The Passive Lysis Buffer (Renilla) is compatible with the Firefly Assay, but the Firefly Lysis Buffer is not compatible with the Renilla Assay.

Category: Luciferase Assays

← FAQs

Luminescence values are relative and depend on the experimental system. For example, luminometers from two different manufacturers each may have 6-log capability, but one may measure a range of 1 to 1,000,000 relative light units (RLU) while the other measures 0.01 to 10,000 RLU. Both instruments can measure a dynamic range of 6-log units, but they read out different arbitrary units.

Category: Luciferase Assays

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← FAQs

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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.