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

Description

The Thermo Scientific™ Pierce Renilla-Firefly Luciferase Dual Assay Kit provides the necessary reagents to simultaneously detect intracellular Renilla and Red Firefly luciferase activity in mammalian whole cell lysates.

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)

Includes:

Cell lysis buffer, reaction buffer, and substrates coelenterazine and D-luciferin

Requires:

Green Renilla and Red Firefly luciferase reporters; Filter sets at 525nm±20nm for Green Renilla Luc and 615 to 675nm for Red Firefly Luc. Luminometer or other instrument capable of monitoring luminescence, such as Thermo Scientific Luminoskan Ascent and Varioskan Flash Microplate Readers; Injector required for assessing more than 24 wells at a time.

Recommended for:

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)