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Firefly Luciferase Assay Kit 2.0

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.

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Product 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. Firefly luciferase is a 62,000 Dalton protein, which is active as a monomer and does not require subsequent processing for its activity. The enzyme catalyzes ATP-dependent D-luciferin oxidation to oxyluciferin, producing light emission centered at 560 nm. Firefly luciferase follows Michaelis-Menten kinetics and, as a result, maximum light output is not achieved until the substrate and co-factors are present in large excess. When assayed under these conditions, light emitted from the reaction is directly proportional to the number of luciferase enzyme molecules. This firefly luciferase assay kit is designed for simple and efficient quantitation of firefly luciferase reporter enzyme activity from cultured cells with high sensitivity and linearity. This kit is a new and improved replacement for the Firefly Luciferase Assay Kit (catalog no. 30003). It features Firefly Assay Buffer 2.0, which has better working solution stability compared to the original assay buffer.

Note: This is a flash-type luminescence assay that requires signal to be measured immediately after adding working solution to samples. It requires a single tube luminometer or luminescence microplate reader with reagent injectors. Biotium also offers the Steady-Luc HTS Firefly Assay Kit (cat. no. 30028L), which is a homogenous glow-type assay with signal half-life of 3-5 hours.

Biotium also offers the Firefly & Renilla Luciferase Single Tube Assay Kit (catalog no. 30081), a combined luciferase assay kit for sequential measurement of Firefly and Renilla luciferase activity sequentially in the same sample with high sensitivity and linearity.

Assay size is based on 96-well format. The kits include enough cell lysis buffer to perform the stated number of assays in plate sizes ranging from 96-well to 24-well. Additional 5X Firefly Lysis Buffer (catalog no. 99923) can be purchased separately if needed.

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. Reporter genes: Application to the study of mammalian gene transcription. Anal. Biochem. 188:245-254.
2. Bronstein, I., et al. 1994. Chemiluminescent and bioluminescent reporter gene assays. Anal. Biochem. 219:169-181.
3. Gould, S.J. and S. Subramani. 1988. Firefly luciferase as a tool in molecular and cell biology. Anal. Biochem. 175:5-13.
4. Brasier, A.R., et al. 1989. Optimized use of the Firefly luciferase assay as a reporter gene in mammalian cell lines. BioTechniques. 7:1116-1122.

 

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