Thermo Fisher, 16185, Pierce™ Renilla-Firefly Luciferase Dual Assay Kit

100 Reaction Kit The Thermo Scientific PierceRenilla-Firefly Luciferase Dual Assay Kit provides the necessary reagents to simultaneously detect intracellularRenillaand Red Firefly luciferase activity in mammalian whole cell lysates.
Features of theRenilla-Firefly Luciferase Dual Assay Kit:
•Simultaneous—concurrent, filter-based, wavelength-separated detection of two luciferase activities
•Sensitive—measure GreenRenillaand 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)The Pierce Renilla-Firefly Luciferase Dual Assay Kit is a highly sensitive assay that allows for the simultaneous detection of GreenRenillaand Red Firefly luciferase activity. GreenRenillaluciferase acts as an experimental reporter with constitutively active red firefly as a normalization control. As shown in the animation video below, 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.
Includes:Cell lysis buffer, reaction buffer, and substrates coelenterazine and D-luciferinRequires:GreenRenillaand Red Firefly luciferase reporters; Filter sets at 525nm±20nm for GreenRenillaLuc 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.
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)GreenRenillaproduces brighter bioluminescent signal than firefly and nativeRenillaluciferases. The bioluminescent signal (λmax= 535nm) produced by GreenRenillaluciferase protein results from the oxidation of coelenterazine. The second assay uses red firefly luciferase, which is a mutant form of the Japanese firefly luciferaseLuciola cruciata. This luciferase produces a red-shifted emission spectrum (λmax= 613nm) that results from the oxidation of D-luciferin in the presence of ATP.
More Product Data
•Simultaneous dual-emission detection of luciferase reporter assaysAssay: Reporter Enzyme, Luciferase Reporter Assay
Compatible Cells: Mammalian Cells
Detection Method: Bioluminescence
For Use With (Equipment): Luminometer (Microplate)
Format: 384-well plate, 96-well plate
Label Type: Enzyme Labeled
Product Line: Pierce™
Quantity: 100 Reaction Kit
Substrate: Coelenterazine, D-Luciferin
Substrate Properties: Chemical Substrate
Substrate Type: Luciferase Substrate
Sufficient For: 100 Dual Luciferase Assays
Target: Luciferase, Firefly Luciferase, Green Renilla Luciferase
Technique: Enhanced Chemiluminescence
Type: Assay Kit
Unit Size: Each