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
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.