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

The Pierce Renilla-Firefly Luciferase Dual Assay Kit is a highly sensitive assay that allows for the simultaneous detection of Green Renilla and Red Firefly luciferase activity. Green Renilla 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 Green Renilla 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

Green Renilla and Red Firefly luciferase reporters; filter sets at 525 nm±20 nm for Green Renilla 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)