Reporter Gene Assays

Reporter gene assays utilizing the luciferase gene, derived from the North American firefly Photinus pyralis, have long been used as a sensitive method to monitor changes in gene expression. Firefly luciferase catalyzes the oxidation of the substrate D-luciferin to produce light, allowing direct measurement of luciferase activity levels via luminescent detection. Thus, expression levels of a gene of interest can be tracked through creation of a recombinant gene fusion with luciferase and subsequent quantitation of luciferase activity.

One useful application of this technology is in high throughput compound screens designed to assay effects on signaling pathway activity. For example, agonist-induced GPCR activation leads to the production of second messengers such as cAMP, IP3 and Ca2+. Second messengers can in turn bind to specific cytosolic proteins known as response element binding proteins. CREB is a representative example of this family of proteins. Upon cAMP binding, CREB translocates into the nucleus and binds to specific DNA sequences (response elements) to initiate gene expression. Re-engineering DNA response elements known to bind CREB, by adding a DNA sequence coding for luciferase, allows one to create a reporter gene system sensitive to cAMP. Such a reporter gene system allows measurement of GPCR activity through luminescence generated by luciferase.

PerkinElmer has developed a suite of proprietary luciferase detection systems which provide superior performance in an easy-to-use format, regardless of the specifics of your experimental design. All of our Lites series of luciferase detections systems offer the following benefits:

• Convenient One-Step Processing: No washes required – simply mix, incubate and read.
• Enhanced Stability: All lites reagents are designed for extended storage at 2-8°C.
• No DTT: Unlike most luciferase detection reagents, lites products do not contain DTT, eliminating toxic hazards and the need for hood work.
• Excellent Z’ Values: All lites products offer robust, sensitive performance, with high signal-to-background ratios and excellent Z’values.

PerkinElmer offers a selection of luciferase detection systems designed to meet all of your reporter gene assay needs. Whether your assay requires the sensitivity provided by high signal intensity, or the flexibility of an extended signal half-life, PerkinElmer provides a solution. Choose from one of our three luciferase detection systems depending on the specific requirements of your experiment.

• neolite™ for:



• Strong signal intensity with an extended signal half-life
• High throughput applications where transfection efficiencies are low
• Screening applications requiring increased assay windows.



• steadylite™ plus for:



• Longest signal half-life
• High throughput screens with extended read times
• Batch processing methodologies



• britelite™ plus for:



• Strongest signal intensity
• Maximum sensitivity
• Continuous processing methodologies