britelite™ plus Reporter Gene Assays


PerkinElmer, Inc.

britelite™ plus offers the same reliability and performance of our current britelite product portfolio. Increased stability, enhanced storage conditions, and odor-free reagents make this new product ...read more

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Description

 

britelite™ plus offers the same reliability and performance of our current britelite product portfolio. Increased stability, enhanced storage conditions, and odor-free reagents make this new product the perfect choice for reporter gene assays.

Luciferase from the North American firefly (Photinus pyralis) is the enzyme of choice for reporter gene assays. Luciferase catalyzes the oxidation of the firefly specific substrate, D-luciferin, to produce light. This reaction is extremely efficient and the quantum yield is the highest of any characterized bioluminescent reaction. The bright signal makes this a valuable enzyme to use for reporting promoter activity.

Light production results from the reaction of the active site (carboxyl group) of D-Luciferin with ATP. This initial reaction leads to the reversible formation of denylluciferin and pyrophosphate. The adenyl-luciferin reacts with molecular oxygen to emit light and produce adenyl-oxyluciferin.

Although extremely sensitive, a serious limitation associated with this technology has been imposed by the very short half-life of the light emission. Flash-type kinetics, result from suicidal adenyloxyluciferin formation at the enzyme surface4. Several substances have been described that may influence this reaction in different ways, or that may enhance the light yield.

The luclite® and britelite assay systems are proprietary mixtures of substances that modify the enzymatic reaction to produce a longer lasting signal by preventing the formation of the adenyl-oxyluciferin at the enzyme surface in a way that is different from CoA-enzyme surface interactions which have been previously described. The differences between luclite® and britelite are the sensitivity and the half-life of the light emitting reaction. The reaction kinetics can be directed to either lower sensitivity and longer half-life or to higher sensitivity and shorter half-life. For britelite plus the higher sensitivity and shorter half-life option has been chosen.

The britelite plus assay system offers the following benefits:

Very high assay sensitivity
Designed for continuous process systems
Suitable for higher density microplates such as 384-well and 1536-well plates
Ideal for smaller-scale sample analysis when maximum sensitivity is needed
Odor free
Convenient storage conditions (2 - 8 °C)

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