One-Step Luciferase Assay System

The One-Step Luciferase Assay System is designed to be used for high-throughput, sensitive quantitation of firefly luciferase activity in mammalian cell culture. The reagent consists of two components, a Luciferase Reagent Buffer (Component A) and Lucifer

Luciferase is the general term given to a class of oxidative enzymes that catalyze reactions that give off light, a process known as bioluminescence (Fig. 1). In biology, researchers can take advantage of this reaction and use it as a readout for various biological processes. This has perhaps been exploited most in luciferase reporter cell lines where a promoter region from a gene of interest is placed immediately upstream of the coding sequence for luciferase. In this system, transcriptional activation of the gene of interest leads to a level of luciferase expression that is proportional to the level of gene activation. The level of activation is then assessed by lysing the cells and adding the luciferase substrate, D-luciferin. Using the One-Step Luciferase Assay System from West Bioscience, this is accomplished in a single step as the lysis buffer has been formulated with D-luciferin. After a brief incubation period, bioluminescence is read on a luminometer.

Luciferin, Firefly luciferase, luciferase buffer

See product datasheet for detailed protocol.

At least 6 months when stored as directed. Upon first thaw, store in aliquots at -20

• The reagent has been validated in a 96-well format. Other formats will require scaling and optimization by the end-user. • Luciferase Reagent Buffer must be at ~ room temperature (20- 25°C) before use. • Luciferase Assay Reagent should be added to cells for at least 5 minutes before measuring luminescence to allow complete cell lysis. • For maximal light intensity, measure samples within 1 hour of reagent addition. • Avoid exposing to excessive heat or light during incubation. • Different cell lines may exhibit variation in lysis ability and/or luminescence signal and may require slight optimization by the end-user. • Luminescence signal is affected by assay conditions. Results should be compared between samples measured using the same cell type and media/serum combinations. • To analyze multiple plates, include a common control sample in each plate and normalize the luminescence of each plate to the control contained in the same plate. • Background luminescence is a characteristic of luminometer performance, therefore, background luminescence must be subtracted from all readings for accuracy.  

Protect from light. Avoid freeze/thaw cycles.