Dual Luciferase Reporter Gene Assay Kit

Reporter gene assays are an important tool in modern molecular biology research for analyzing the interaction between potential cis-elements (such as promoters, enhancers, and silencers) and trans-acting factors in the flanking regions of structural genes. The Dual Luciferase Reporter Gene Assay Kit is an advanced auxiliary reporter gene technology combining firefly and Renilla luciferase detection.

Although both firefly and Renilla luciferases have the characteristics of bioluminescent reporter genes, they originate from entirely different evolutionary backgrounds, and thus have completely different enzyme structures and substrate requirements. We exploit this difference by using both enzymes together, forming a highly effective dual luciferase reporter gene system, with Renilla luciferase typically serving as an internal control.

The experimental principle involves using firefly luciferin as the substrate to detect the firefly luciferase in cells. Firefly luciferase catalyzes the oxidation of luciferin in the presence of ATP, Mg2+, and O2, producing oxyluciferin and generating bioluminescence during the oxidation of luciferin. In subsequent steps, the reaction is inhibited to stop the bioluminescence from the firefly luciferin, and the substrate coelenterazine is added for Renilla luciferase. In the presence of O2, Renilla luciferase catalyzes the oxidation of coelenterazine to coelenteramide, generating bioluminescence. The bioluminescence is then measured using a chemiluminescence or scintillation counter.

The dual luciferase reporter gene assay system introduces an "internal control" reporter gene in addition to the single luciferase reporter gene. This reduces the impact of external factors such as cell number, cell growth state, and transfection efficiency on the experimental results, making the data more reliable.

Features:
· Quick and simple
· High sensitivity
· Safe and reliable
· Stable and accurate

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