Thermo Scientific™ Pierce™ Gaussia-Firefly Luciferase Dual Assay Kit

The Pierce Gaussia-Firefly Luciferase Dual Assay Kit is a highly sensitive assay that allows for the simultaneous detection of Gaussia and Red Firefly luciferase activity. Gaussia luciferase acts as an experimental reporter with constitutively active red firefly as a normalization control. This reporter-and-control combination enables simultaneous monitoring of experimental reporter and control luciferase activities in a single-read assay without the need for two-step addition of substrate reagents or quenching.

The assay working solution contains substrates for both luciferases, and the reactions occur simultaneously with flash-type kinetics. The resulting luminescent signals are spectrally resolvable using filters. In a single sample, researchers can assay transcriptional activity of regulatory elements, signal transduction pathways, and effects of activators or inhibitors.

Highlights

• Simultaneous—concurrent, filter-based, wavelength-separated detection of two luciferase activities
• Sensitive—measure bright blue Gaussia and red firefly luciferase activities in the same sample
• Fast—no quenching step required, unlike in traditional sequential dual assays
• Multiplex—capable of quantitating two cellular activities in the same sample(s)

Requires

Gaussia and Red Firefly luciferase reporters; filter sets in the ranges of 425 to 525 nm for Gaussia Luc and 615 to 675 nm for Red Firefly Luc; luminometer or other instrument capable of monitoring luminescence, such as the Varioskan ALF or the Varioskan LUX microplate readers; injector for assessing more than 24 wells at a time

Recommended applications

Study two regulatory elements at the same time; monitor two signaling pathways simultaneously; enable studying more than one target per screen (e.g., off-target effects)