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 Form liquid Luciferase Firefly and Renilla Signal Half-life 30 min Number of Product Components 5 Workflow Two Step Components Components No. Name 11402ES60 11402ES80 11402-A Cell Lysates 20 mL 10×20 mL 11402-B Firefly Luciferase Buffer 10 mL 10×10 mL 11402-C Firefly luciferase substrate (50 ×) 200 μL 10×200 μL 11402-D Renilla Luciferase Buffer 10 mL 10×10 mL 11402-E Renilla Luciferase substrate (50 ×) 200 μL 10×200 μL Figures 1. Product Performance Is Consistent with Imported Products Figure 1 shows the comparison of Firefly luciferase luminescence intensity, quenching efficiency, and Renilla luciferase luminescence intensity. Yeasen 11402 Dual-Luciferase Reporter Assay product performs comparably to Brand P in terms of luminescence intensity. Meanwhile, Yeasen 11402 is also on par with Brand P in Firefly luciferase quenching efficiency. Additionally, the residual fluorescence of Yeasen 11402 is extremely low and negligible, ensuring no cross-interference with subsequent Renilla luciferase signal generation, thereby guaranteeing the accuracy and reliability of experimental results. 2. Superior Signal Stability Figure 2 shows the kinetic testing results for the Firefly luciferase luminescence signal stability of Yeasen 11402 Dual-Luciferase Reporter Assay product. Tracking the Firefly luciferase luminescence signal continuously for up to 30 minutes from the start of the reaction revealed almost no signal decay in Yeasen 11402. This exceptional signal stability far exceeds the time required for standard microplate reader operations, fully meeting practical detection requirements. 3. Five Freeze-Thaw Cycles Have Minimal Impact on Signal Figure 3 displays the luminescence signals of Firefly luciferase and Renilla luciferase after subjecting the Yeasen 11402 Dual-Luciferase Reporter Assay product to 5 freeze-thaw cycles. Results indicate that signal intensity remains virtually unaffected with extremely high stability. This feature enhances its practicality in laboratory settings to better satisfy daily operational needs.
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