Firefly luciferase is a protein with a molecular weight of about 61 kDa, which can catalyze the oxidation of luciferin to oxyluciferin in the presence of ATP, magnesium ions and oxygen. It emits bioluminescence with a wavelength of around 560 nm. Renilla luciferase is a protein with a molecular weight of about 36 kDa, which can catalyze the oxidation of coelenterazine to coelenteramide in the presence of oxygen, and emits a wavelength of 480 nm during the oxidation process. about bioluminescence. Both bioluminescence can be measured by chemiluminescence. The detection principle is shown in the figure 1:

 Figure 1: Firefly and Renilla luciferase assay schematic

Usually, the 5´UTR or promoter of the target gene is cloned upstream of Firefly Luciferase, or the 3´UTR is cloned downstream of Firefly Luciferase, and the transcriptional regulation effect of the promoter or regulatory element is detected by detecting the amount of firefly luciferase. Renilla Luciferase was used as an internal control to eliminate differences in cell number, transfection efficiency, etc. The Dual Luciferase Reporter Gene Assay Kit uses luciferin as a substrate to detect the activity of the firefly luciferase reporter gene, and then uses coelenterazine as a substrate to detect Renilla luciferin while quenching the fluorescence reaction. Enzyme reporter gene activity. The kit has the characteristics of high sensitivity.

Features

Enhanced Lysis Capability: Capable of completely lysing the vast majority of cell types.

Stronger Signal: Precise detection of weak promoter expression.

Higher Sensitivity: Detectable down to 10-20 picomoles of luciferase molecules.

Broader Linear Range: Linear detection range exceeds 8 orders of magnitude of enzyme concentration.

Applications

 Verification of miRNA interactions with target gene 3'UTRs

Verification of lncRNA-miRNA interactions

Verification of transcription factor regulation in the promoter region

Signaling pathway analysis

SNP activity analysis

Specifications

Components

Shipping and Storage

The Dual Luciferase Reporter Gene Assay Kit products should be stored at -15℃ ~ -25℃ for 1 year.

Figures

Figure1. YEASEN (Cat#: 11402) has a fluorescence intensity comparable to that of Brand P.

Figure2. YEASEN (Cat#: 11402) detection reagents interact with luciferase to achieve optimal luminescence intensity. After the addition of Renilla luciferase substrate, the bioluminescence of firefly luciferase is quenched: it can be essentially completely quenched.                                                           

Documents:

Safety Data Sheet

11402 MSDS HB220209 EN.PDF

Manuals

11402 Manual HB241230 EN.pdf

Citations & References: 

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