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Application: Monitor firefly (Photinus pyralis) luciferase activity in cultured mammalian cells. High-throughput screening using luciferase reporter cell lines
Background: 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 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.
Description: 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 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.
Storage Stability: At least 6 months when stored as directed. Upon first thaw, store in aliquots at -20°C. Thereagent may be subjected to several freeze/thaw cycles with no effect on functionality, but it isrecommended that freeze/thaw cycles be avoided whenever possible.
Warnings: Protect from light. Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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