The ONE-Step™ Luciferase Assay System is designed 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 reagents for cell lysis and luciferase quantitation.
Sensitive – highly sensitive detection of firefly luciferase activity.
Stable – the signal is stable for more than two hours, providing flexibility regarding incubation time.
Convenient – simple one-step, homogeneous protocol.
High-throughput – the one-step 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 which catalyze reactions that give off light, a process known as bioluminescence (Figure 1). Enzymatic activity is directly proportional to the expression of luciferase in the cells and can be quantified upon addition of the appropriate substrate.
Figure 1. Bioluminescent reaction catalyzed by luciferase.
In the luciferase reaction, the cells are lysed with luciferase substrate containing D-luciferin. Firefly luciferase expressed in reporter cells catalyzes the conversion of D-luciferin to oxyluciferin using ATP as a co-substrate, and in the presence of Mg2+, in a reaction that gives off light. The amount of light is proportional to the amount of luciferase present in the reaction. Luminescence is read using a luminometer.
Luciferase is the general term given to a class of oxidative enzymes that catalyze reactions that give off light, a process known as bio-luminescence (Fig.
Gene reporter assays are widely used in research and Drug Discovery. The development of such cell lines is often time-consuming and costly, sometimes upwards of
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