Dual-Light® Combined Reporter Gene Assay System for Detection of Luciferase and beta-GalactosidaseApplied Biosystems
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.
- Detection of firefly luciferase and b-galactosidase in the same sample simplifies normalization of transfection efficiency
- Wide dynamic range of b-galactosidase and luciferase assays enables accurate measurement of both enzymes from femtogram to nanogram range
- Assay sensitivity is 100- to 1,000-fold better than either the isotopic/non-isotopic assays for chloramphenicol acetyl transferase (CAT) or the colorimetric/fluorescent assays for b-galactosidase, providing greater sensitivity than competing assay technologies
- Highly sensitive assay with a wide dynamic range permits detection of high and low levels of reporter without performing numerous sample dilutions
- Non-radioactive reporter gene assay kit eliminates concerns over use of radioisotopes
- Assay can be completed in about one hour, providing fast assay turnaround
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.