What is firefly luciferase assay?
The Firefly Luciferase HTS Assay is a steady-glow high sensitivity firefly luciferase reporter gene assay kit for the quantification of firefly luciferase expression in mammalian cells with signal half-life of about 3 hours.
How does a luciferase assay work?
A Luciferase Reporter Assay. When this protein activates transcription, the cell will produce luciferase enzyme. After the addition of a lysis buffer and a substrate, a luminometer quantifies the luciferase activity. If your protein activates the expression of the target gene, the amount of signal produced increases.
What is the purpose of a dual luciferase assay?
Allows study of weak promoters, low-level expression/regulation and expression in cells that transfect poorly.
What does luciferase do in firefly?
Firefly luciferase catalyzes the oxidation of firefly luciferin, producing a short-lived flash of light that decays within a few seconds.
How is luciferase made in fireflies?
This type of light production is called bioluminescence. The method by which fireflies produce light is perhaps the best known example of bioluminescence. When oxygen combines with calcium, adenosine triphosphate (ATP) and the chemical luciferin in the presence of luciferase, a bioluminescent enzyme, light is produced.
Where is luciferase produced in a firefly?
The amino acid sequences on the surface of the two domains facing each other are conserved in bacterial and firefly luciferase, thereby strongly suggesting that the active site is located in the cleft between the domains.
How do reporter gene assays work?
Reporter gene assays are typically used to measure the regulatory ability of an unknown DNA-sequence. This is done by linking the unknown promoter sequence to an easily detectable reporter gene whose product can be easily detected and quantifiably measured.
How does Gaussia luciferase work?
Gaussia luciferase is a 20kDa protein from the marine copepod, Gaussia princeps. The bioluminescent enzyme is highly secreted into the cell culture media, allowing for live cell monitoring of reporter activity.