The 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 Luciferase Reagent Substrate (Component B). Component A and Component B are combined to form a working solution that contains all the necessary components for cell lysis and luciferase quantitation. This assay system has several features: • Sensitive-highly sensitive detection of firefly luciferase activity. • Stable-the signal output is stable for more than two hours, providing flexibility with regard to incubation time • Convenient-simple one-step, homogeneous protocol. • High-throughput-one-step homogeneous protocol minimizes handling steps to support high-throughput screening applications • Compatibility-works well with a variety of common media containing 0-10% serum and phenol red. • Instrumentation-does not require a luminometer with injectors.
Luciferase is the general term given to a class of oxidative enzymes that catalyze reactions that give off light, a process known as bioluminescence. 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 BPS 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.
Monitor firefly (Photinus pyralis) luciferase activity in cultured mammalian cells. • High-throughput screening using luciferase reporter cell lines
At least 6 months when stored as directed. Upon first thaw, store in aliquots at -20°C. The reagent may be subjected to several freeze/thaw cycles with no effect on functionality, but it is recommended that freeze/thaw cycles be avoided whenever possible.
Luciferase Reagent Buffer (Component A)Amount:2 x 50 ml Storage: -20°C Luciferase Reagent Substrate, 100x (Component B) Amount: 2 x 500 µl Storage: -20°C, Protect from light Each system contains sufficient reagents to perform 1,000 assays of 100 μL each in 96-well plate.