Dual-Light® luminescent reporter gene assay is designed for the rapid and sensitive detection of firefly luciferase and b-galactosidase in a single extract aliquot for gene expression assays.
Light signal from each enzymatic reaction is measured sequentially in a luminometer with automatic injectors or other instrumentation in which light emission measurements can be performed within a short period. First, luciferase reporter enzyme activity is quantitated with an enhanced luciferase reaction. Following a 30-60 minute incubation and addition of a light-emission accelerator, b-galactosidase reporter enzyme activity is determined with Galacton-Plus® substrate. Both assays are combined into one simple assay using only one extract aliquot for greater convenience and precision. The entire assay is completed in less than one hour.
Wide Dynamic Range
The wide dynamic range of this dual assay enables accurate measurement of luciferase and b-galactosidase concentrations over seven orders of magnitude, from the femtogram to nanogram range. Colorimetric and isotopic reporter gene assays cannot rival the dynamic range of the Dual-Light® system. We have formatted this assay for tube or microplate luminometer, with injection capability.
Dual-Light reporter gene assay system has been very widely used for reporter quantitation/transfection normalization from transiently transfected mammalian cell lines as well as transfected primary cells. In addition, it has been used with a modified lysis buffer to quantitate luciferase and beta-galactosidase activities from a novel reporter fusion construct in yeast cells. For these and additional references, please see our bibliography in the Literature/Resources section.