What is a dual luciferase assay?
What is a dual luciferase assay?
The dual luciferase assay is performed by sequentially measuring the firefly and Renilla luciferase activities of the same sample, with the results expressed as the ratio of firefly to Renilla luciferase activity (Fluc/Rluc).
What is the purpose of a dual luciferase assay?
The Dual Luciferase (Firefly-Renilla) Assay System is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture. The Firefly Luciferase Reagent is first added to the cells in medium directly.
What is firefly luciferase assay?
The Firefly/Renilla Dual Luciferase Assay (SCT152) allows measurement of both Firefly and Renilla luciferase activity in the same sample with high sensitivity and linearity. This is a flash-type assay that requires luminescence to be measured immediately after adding the detection reagents to the luciferase sample.
What is Renilla luciferase?
Renilla luciferase enzyme/protein (RL), purified from sea pansy (Renilla reniformis), is a bioluminescent soft coral that displays blue-green bioluminescence upon mechanical stimulation. The enzyme RL catalyzes coelenterazine oxidation leading to bioluminescence.
How do you do dual luciferase assay?
Sensitive, Rapid and Convenient Dual-Reporter Assay After quantifying the firefly luminescence, this reaction is quenched, and the Renilla luciferase reaction is initiated simultaneously by adding Stop & Glo® Reagent to the same sample.
What does a luciferase assay tell you?
A luciferase assay is used to determine if a protein can activate or repress the expression of a target gene. Luciferase is an enzyme used for bioluminescence by various organisms in nature, most famously the firefly.
Why is luciferase the best reporter?
Luciferase-based assays are better than other reports because of the following advantages: Quick and real-time measurement. Exceptionally high sensitivity than fluorescent reporters like GFP (10- to 1,000 fold) Range of measurement is wide and dynamic.
How do reporter assays work?
To perform the reporter assay, you clone the regulatory region of your gene-of-interest (X) upstream of the luciferase gene in one of these expression vectors, introduce that resulting vector DNA into cells, and let the cells grow for a period of time to allow transcription and translation to occur.
Where is firefly luciferin found?
Lampyridae
Firefly luciferin is the luciferin found in many Lampyridae species. It is the substrate of beetle luciferases (EC 1.13. 12.7) responsible for the characteristic yellow light emission from fireflies, though can cross-react to produce light with related enzymes from non-luminous species.
What is NanoLuc?
NanoLuc™ (Nluc) is a 19.1 kDa, ATP- independent luciferase that utilizes a novel coelenterazine analog (furimazine) to produce high intensity, glow-type luminescence.
How does Gaussia luciferase work?
Gaussia luciferase can be expressed in mammalian cells using commercially available reporter plasmids. This luciferase, which does not require ATP, catalyzes the oxidation of the substrate coelenterazine in a reaction that emits light (at 470 nm), and has considerable advantages over other reporter genes.
