The differences between single and dual luciferase detection applications

Reporter gene assays are primarily divided into single-luciferase reporter gene assays and dual-luciferase reporter gene assays. In specific applications, how can you determine whether experiment uses a single-luciferase or dual-luciferase detection system? Let's take a look at the differences.

Single Luciferase Reporter Assay System

Number of Reporter Genes: A single firefly luciferase reporter gene;

Transfection Type: A single reporter gene plasmid, after transfection, drug selection is used to construct a stable reporter gene cell line expressing luciferase;

Detection Process: Only the firefly luciferase signal needs to be detected, simple operation, short detection period, suitable for high-throughput drug screening;

Application Scenario Analysis: It is usually through the construction of a stable overexpressing monoclonal cell line, as the expression level of these cell lines is relatively stable and uniform, without the influence of experimental variations due to transfection efficiency and cell viability. The operation is simple, and since only the firefly luciferase signal is detected, data analysis is more convenient. Coupled with the advantages of short detection time, it is particularly suitable for high-throughput drug screening and meets the needs of large-scale sample detection.

Dual-Luciferase Reporter Gene Detection System

Reported Gene Quantity: Dual reporter genes of firefly luciferase and Renilla luciferase;

Transfection Type: Co-transfection of two reporter gene plasmids, mainly through transient transfection;

Detection Process: It is necessary to first detect the firefly signal and then the Renilla signal, and the ratio of the two (Firefly/Renilla) is used for the final data calculation;

Application Scenario Analysis: It is generally through transient transfection, with the co-transfection of a plasmid containing the Renilla luciferase gene as an internal reference, providing a benchmark for the experiment, thereby reducing the interference of external factors such as cell viability and transfection efficiency. Through ratio calculation, it is possible to correct for variations in cell number, transfection efficiency, and cell activity, improving the accuracy of experimental results. It is an important tool in the fields of gene expression regulation, signal transduction pathways, and drug screening.

product series

Yeasen Luciferase Reporter Gene Detection product series provides a convenient and efficient quantitative detection solution. The single-component design simplifies the usage process, and the perfect combination of high sensitivity with a super long signal half-life of 5 hours meets the needs of large-scale drug screening detection. We offer three products: Bio-One Step, Steady-One Step, and Bright-One Step, to adapt to different sensitivity and stability requirements.

Product Advantages

Long signal half-life: The signal half-life can reach up to 5 hours, meeting the needs of high-throughput experiments and avoiding signal differences caused by the time difference between the first and last wells due to a large number of samples, ensuring more stable and reliable results.

Strong fluorescence signal: The signal value can reach 8 orders of magnitude, providing a large detection window and strong data support for early drug screening and efficacy evaluation.

Easy operation: One-step operation avoids complicated experimental procedures. Simply add the detection reagent and wait for 5 minutes before starting the machine for detection.

Multiple choices: According to different application scenarios, there are 3 different products available, which can be selected based on the experimental requirements for signal value and signal stability.

Product Data

01  Comparison of Performance for Yeasen Different Reporter Gene Assay Kits

The verification of three types of Assay Kits was conducted on cell lines stably expressing firefly luciferase, and the results showed that the Bright-One Step product has higher signal values; the Steady-One Step product has a longer signal half-life and more stable signals; the Bio-One Step product not only has high signal values but also maintains a longer signal half-life.

Figure 1. Comparison of Performance for Different Yeasen Reporter Gene Assay Kits

02  Yeasen Bio-One Step Luciferase Reporter Gene Detection Reagent Comparison with Competing Products

When using Yeasen Bio-One Step, competitor P, and competitor V on a cell line that stably overexpresses firefly luciferase, the results show that the signal value and signal half-life of Yeasen Bio-One Step are essentially consistent with competitor P, and the signal half-life is superior to competitor V.

Figure 2. Comparison of Yeasen Bio-One Step Luciferase Reporter Gene Detection Reagent with Competitors

03  Yeasen Bio-One Step Luciferase Reporter Gene Detection Reagent Freeze-Thaw Stability Test

Yeasen Bio-One Step Luciferase Reporter Gene Detection Reagent has undergone repeated freeze-thaw experiments, and the results show that even after 5 freeze-thaw cycles, the product performance remains stable with no significant changes.

Figure 3. Repeated Freeze-Thaw Test of Yeasen Bio-One Step Luciferase Reporter Gene Detection Reagent

Operational Procedure

01 Cell Transfection

Directly add the reagent to the cell suspension, then pipette up and down 5 times to ensure thorough mixing, wait for 5 minutes to allow for complete cell lysis.

02 Sample Loading

Pipette 100 μL of the mixture (detection reagent + cell culture medium) into each well of the detection plate.

03 Fluorescence Detection

Set the parameters on the plate reader. Place the detection plate into the plate reader, vibrate it for a few seconds, and then proceed with the detection.

Product List