Assay Kit Mechanism

Reporter gene assays are an important tool for monitoring cellular events associated with gene expression, gene regulation, and signal transduction. Firefly luciferase, one of the most commonly used reporters, produces light by catalyzing a bioluminescence reaction that oxidizes D-luciferin in the presence of oxygen and ATP. Renilla luciferase catalyzes coelenterazine oxidation to produce light. Due to the difference in substrate requirements, it is possible to measure the activity of both luciferases in a single sample through sequential addition of the substrates. The result is a convenient assay with two readouts that can be normalized to each other to provide more comprehensive information about a cellular pathway or event.