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Luciferase Assay Kit

Components:   

10x Cell Lysis Solution: 25 ml, store at 4ºC

Luciferase Substrate: 20 ml (reconstituted in buffer; for 200 assays), store in aliquots at -75ºC

Product Description: Luciferase encoded by the firefly luciferase gene is widely used as a sensitive reporter enzyme for the study of transcriptional regulation. The enzyme catalyzes, in the presence of ATP (see ATP Assay Kit), the oxidation of luciferin with concomitant emission of yellow-green light, which can be conveniently measured by scintillation counters or luminometers. Light emission peaks in several seconds at 560 nm when the reaction is conducted at pH 7.8 (Anal. Biochem. 80:496,1977). The rapid appearance and decay of the light flash require consistent timing of the light measurement to obtain reliable data. This provides the basis for an assay system many times more sensitive than the b-galactosidase (GAL) or other reporter gene systems (PNAS USA 82:7870,1984 & Mol. Cell. Biol. 7:725,1987). The luciferase-based reporter assay is thus well suited for those cell systems exhibiting low transfection efficiency. The kit is sufficient for 200 assays using 0.1 ml of Luciferase Substrate per assay. Reconstituted Luciferase Substrate solution if stored in aliquots at -75°C is stable for at least one year. Repeated freeze-thaw cycles should be avoided. The product is for research use only.

Features: The assay system is simple, non-radioactive, and extremely sensitive. The Cell Lysis Solution, which should be diluted 20-fold with distilled water prior to use, is compatible with the CAT and GAL assays (see b-Galactosidase Assay Kit and Staining Kit). The assay can be completed in as short as 10 minutes. The method is adaptable to automation.