Dual Luciferase Reporter Gene Assay Kit _ 11402ES
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Dual Luciferase Reporter Gene Assay Kit _ 11402ES
Dual Luciferase Reporter Gene Assay Kit _ 11402ES
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...

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Dual Luciferase Reporter Gene Assay Kit _ 11402ES

YeasenSKU: 11402ES10
Size: 10 T
Price:
$25.00
Stock:
In stock

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.

Storage

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

FAQ

Yes, they can be stored at -80°C, similar to standard protein storage protocols. Lysed samples can be stored at -80°C for up to 6 months and at -20°C for up to 1 month.
Strict light protection is not required during operation. However, store them protected from light. Low temperature is a more critical storage condition—especially for coelenterazine (the Renilla substrate), which is recommended to be stored at -80°C.
The ratio should be adjusted according to specific experimental conditions. Pilot experiments are recommended—for instance, testing Renilla vector to Firefly vector ratios at 1:10, 1:20, 1:50, and 1:100.
They are assayed in the same well. There is no mutual interference because the Renilla luciferase substrate reagent contains a compound that quenches Firefly luminescence. While separate assays are possible, sequential measurement in a single well is standard.
Different microplate readers vary in sensitivity and parameter settings, which can lead to differences in background signal readings for the same sample.
Opaque white microplates.
Add 0.3 mL of Triton X-100 to 10 mL of PBS to prepare a 3% Triton X-100 solution.
Dilute the cell lysate sample 5-fold prior to testing.
1. Luciferase is expressed intracellularly and is a enzyme non-native to humans (yielding extremely low background). It cannot cross cell membranes. In contrast, luciferase substrates are hydrophobic, cyclic small molecules that penetrate cell membranes easily. Thus, any luciferase expression is exogenous, and adding substrate directly to culture media or injecting it in vivo yields detectable signals.
2. In vitro reporter assays require cell lysis to homogenize samples, overcoming transfection heterogeneity, uneven substrate permeability, and signal variation. In vitro reporter assays are highly quantitative and sensitive, requiring ATP and oxygen. Cell lysis fully releases expressed luciferase to optimize the reaction; since intracellular oxygen is limited, omitting lysis leads to signal reduction (some lysis buffers include optimized additives like ATP to boost signal). Therefore, cell lysis ensures superior signal in in vitro assays. 3. In vivo imaging instruments possess high sensitivity and power capable of penetrating skin and tissue, enabling strong signal detection without cell lysis.
No, the order cannot be adjusted. In this kit, Component B (Firefly Luciferase Buffer) is essential for detecting both Firefly and Renilla luciferases. The optimal reaction environment for Renilla luciferase requires the presence of Component B; otherwise, Renilla luciferase readings will be extremely low.

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