Dual Luciferase Reporter Gene Assay Kit

Description

Firefly luciferase is a ~61 kDa protein that catalyzes the oxidation of luciferin in the presence of ATP, magnesium ions, and oxygen, producing bioluminescence with an emission peak at approximately 560 nm. Renilla luciferase is a smaller protein (~36 kDa) that catalyzes the oxidation of coelenterazine in the presence of oxygen, emitting light at around 480 nm. The bioluminescent signals from both enzymes can be quantitatively measured using a chemiluminescence analyzer.

In dual-luciferase reporter assays, the promoter or 5′ untranslated region (UTR) of a target gene is typically cloned upstream of the firefly luciferase gene, or the 3′ UTR is cloned downstream, allowing transcriptional or post-transcriptional regulation to be evaluated based on firefly luciferase activity. Renilla luciferase is co-expressed as an internal control to normalize variations caused by differences in cell number and transfection efficiency. The Dual Luciferase Reporter Gene Assay Kit sequentially measures firefly luciferase activity using luciferin, followed by Renilla luciferase activity using coelenterazine after quenching the first reaction, providing high sensitivity and reliable results.