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| Catalog Number | Quantity |
|---|---|
| 16186 | 1000 Reaction Kit |
| 16185 | 100 Reaction Kit |
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.
Features of the 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. 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-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 the Varioskan ALF or the Varioskan LUX microplate readers; Injector required for assessing more than 24 wells at a time.
Applications:
Green Renilla produces brighter bioluminescent signal than firefly and native Renilla luciferases. The bioluminescent signal (λmax= 535nm) produced by Green Renilla luciferase protein results from the oxidation of coelenterazine. The second assay uses red firefly luciferase, which is a mutant form of the Japanese firefly luciferase Luciola cruciata. This luciferase produces a red-shifted emission spectrum (λmax= 613nm) that results from the oxidation of D-luciferin in the presence of ATP.
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