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Sigma-Aldrich

Firefly/Renilla Dual Luciferase Assay

Flash-type dual luciferase assay that allows measurement of both Firefly and Renilla luciferase activity in the same sample with high sensitivity and linearity.

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Synonym(s):
Firefly Luciferase Assay, Dual Luciferase Assay, Bioluminescent Assay

technique(s)

cell based assay: suitable

detection method

chemiluminescent

shipped in

ambient

General description

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 Dual Luciferase Assay allows measurement of both 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 simultaneously quench firefly luciferase activity and measure Renilla luciferase activity. 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. This is a flash-type assay that requires luminescence to be measured immediately after adding the detection reagents to the luciferase sample. Firefly signal decays over the course of about 12 minutes, while Renilla signal decays over the course of about 2 minutes, although this may vary depending on enzyme levels.

Application

Firefly Luciferase Assay
Flash-type dual luciferase assay that allows measurement of both Firefly and Renilla luciferase activity in the same sample with high sensitivity and linearity.
Research Category
Cell Signaling
Research Sub Category
Live Cell Dye

Components

1) 1X Passive Luciferase Lysis Buffer 2.0 (CS224514): 15 mL

2) Firefly Luciferase Assay Buffer 2.0 (CS224591): 15 mL

3) D-Luciferin (CS224551): 3 X 1 mg

4) Renilla Luciferase Assay Buffer 2.0 (CS224580): 15 mL

5) Aquaphile Coelenterazine (CS224581): 3 X 200 µg

Physical form

Liquid

Storage and Stability

Store Firefly/Renilla Dual Luciferase Assay at -80°C. Firefly and Renilla Assay Buffers are stable at -80°C for at least six months from date of receipt. Other components are stable at -20°C or below for at least six months from date of receipt. Kit components and stock solutions of D-Luciferin and Aquaphile Coelenterazine in water are stable to at least 5 freeze/thaw cycles.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

Regulatory Information

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Xinyu Tang et al.
Bioengineered, 12(1), 4643-4653 (2021-07-30)
Non-small cell lung cancer (NSCLC) is one of the main causes of death in the world. To improve the diagnostic level and find new biological targets，GSE datasets were selected from GEO databaseto analyze the differential expression genes and construct ceRNA
Joseph M Chan et al.
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Small cell lung cancer (SCLC) is an aggressive malignancy that includes subtypes defined by differential expression of ASCL1, NEUROD1, and POU2F3 (SCLC-A, -N, and -P, respectively). To define the heterogeneity of tumors and their associated microenvironments across subtypes, we sequenced

Articles

Bioluminescent Firefly Luciferase Assays

Firefly luciferase is a widely used bioluminescent reporter for studying gene regulation and function. It is a very sensitive genetic reporter due to the absence of endogenous luciferase activity in mammalian cells or tissues.

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