Firefly Luciferase Reporter Gene Assay Kit
$190.00 - $1,100.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: FLR-100 (for 100T)
Cat. No.: FLR-1k (for 1000T)
Description
Firefly Luciferase Reporter Gene Assay Kit is a reagent kit used to detect the expression of the firefly luciferase reporter gene in ADCC Reporter Bioassays or cells using a chemiluminescence-based method. It offers high sensitivity and signal stability. This assay kit provides flexibility and convenience, along with high detection sensitivity.
This product utilizes a novel substrate for luciferase, which results in a more stable reaction with samples and greater tolerance to sample components compared to standard luciferase analysis reagents. It is less susceptible to the effects of mixing and sample conditions, thereby enhancing the reproducibility of results. It is an ideal choice for ADCC Reporter Bioassays and other firefly luciferase reporter gene activity assays.
Features
- This product exhibits high detection sensitivity and has a wide linear range for sample measurement. In a 96-well plate, typically, this product demonstrates a good linear relationship within a range of 200,000 cells. The upper limit of detectable cells may vary depending on different cell types and levels of firefly luciferase expression. When the expression level of firefly luciferase is very high, there may be a good linear relationship only within a range of 100,000 cells. However, the chemiluminescence readings will still continue to increase with an increase in the number of cells.
- This product is easy to use, provides stable readings, and has a fast detection speed, completing the assay in approximately 10 minutes. It offers a simpler and more convenient alternative compared to other methods for firefly luciferase detection. Cells can be cultured and assayed in the same multiwell plate without the need for cell washing or changing/removing culture media. Simply dissolve the detection substrate (lyophilized powder) in the provided assay buffer to prepare the Firefly Luciferase Assay Reagent. Mix the reagent with the cultured cells or equivalent volume, allow a 5-minute reaction, and then proceed with chemiluminescence detection. The generated chemiluminescent signal is highly stable, retaining over 70% of the signal even after 60 minutes of the reaction. A versatile microplate reader capable of measuring chemiluminescence in a 96-well plate usually completes the detection of one plate within 2 minutes.
- This product exhibits excellent stability. It can be stored long-term at -20°C, and even after 5 freeze-thaw cycles, it shows no significant impact on the detection performance. The detection efficiency decreases by no more than 10% after 10 freeze-thaw cycles. When stored at 4°C, the Firefly Luciferase Assay Reagent retains less than a 20% decrease in detection efficiency after 3 days, less than a 30% decrease after 5 days, and maintains over 65% of the detection efficiency after 7 days. It retains over 70% of the detection efficiency when stored at room temperature for 1 day, over 60% for 3 days at room temperature, and over 60% for 1 day at 37°C. Even when the detection efficiency is reduced to approximately 50%, the Firefly Luciferase Assay Reagent can still meet various routine testing requirements.
- This product offers flexibility and convenience. It is suitable for both small-scale and high-throughput screening of samples. It can be directly added in equal volumes to the cell culture wells for detection or used to detect samples after cell lysis and collection using the RG126S/M Firefly Luciferase Reporter Gene Cell Lysis Buffer.
- This product exhibits strong compatibility. It is compatible with various common culture media and shows minimal impact on signal and stability from phenol red, up to 10% fetal bovine serum or calf serum, up to 2% DMSO or ethanol, and luciferase inhibitors. Commonly used salts and metal ions at normal concentrations also have minimal effect.
Principal
Firefly luciferase is a protein with a molecular weight of approximately 61 kDa. In the presence of ATP, magnesium ions, and oxygen, it catalyzes the oxidation of luciferin to oxyluciferin, emitting bioluminescence at a wavelength of around 560 nm. Bioluminescence can be measured using a luminometer or a liquid scintillation counter. The bioluminescent system of luciferin and luciferase enables sensitive and efficient detection of gene expression. Typically, the regulatory elements or 5' promoter region of the gene of interest are cloned upstream of the luciferase gene, or the 3'-UTR region is cloned downstream of the luciferase gene, creating a reporter gene plasmid. The plasmid is then transfected into cells, and after treatment with appropriate drugs or agents, the cells are lysed, and the luciferase activity is measured. The level of luciferase activity indicates the transcriptional regulation of the target gene by the drug or treatment. In ADCC Reporter Bioassays, ADCC effector cells are genetically modified to stably express the firefly luciferase system driven by NFAT (nuclear factor of activated T-cells) responsive elements and Fc receptors (such as FcγRIIIa receptor). When the Fab fragment of an antibody specifically binds to the corresponding antigenic epitope on target cells, the Fc fragment of the antibody binds to the Fc receptors on effector cells, activating the NFAT signaling pathway and inducing the expression of firefly luciferase. The Firefly Luciferase Reporter Gene Assay Kit is used to quantitatively detect the expression of firefly luciferase by adding the assay reagents and using a luminometer for luminescence detection.
Additional Notes
- Due to the sensitivity of luciferase activity to temperature, both cells and assay reagents should be equilibrated to room temperature before performing the assay. The assay buffer can be thawed and mixed at room temperature or in a water bath not exceeding 25°C before mixing with the substrate to prepare the assay reagent.
- Although it has been tested that repeated freezing and thawing of the assay buffer and substrate mixture for up to 5 cycles does not significantly affect the assay performance, it is recommended to prepare the assay reagent fresh for optimal stability and performance. Any unused assay reagent should be appropriately aliquoted, protected from light, and stored to avoid repeated freezing and thawing as well as prolonged exposure to room temperature. During the freezing and thawing process, a small amount of precipitation may occur in the assay reagent. In such cases, it should be equilibrated to room temperature and dissolved as much as possible. If there are still residual insoluble particles, they can be removed by centrifugation before use, as it has been tested that they do not affect the subsequent assay performance.
- Please use white or black 96-well or 384-well plates suitable for cell culture. Using regular transparent 96-well or 384-well plates may cause interference between adjacent wells.
- High solvent content in the test drug may interfere with the luciferase reaction and affect the chemiluminescent signal. This interference can be eliminated by setting control wells containing cell culture medium with the solvent. It has been tested that a DMSO content of up to 2% in the final reaction system does not affect the reaction.
- This product is for scientific research purposes only and is intended for use by professionals. It should not be used for clinical diagnosis or treatment, food or drug purposes, and should not be stored in regular residential areas.
- For your safety and health, please wear laboratory attire and disposable gloves when handling.
Storage
Store at -20°C in the dark for at least one year. The Firefly Luciferase Reporter Gene Assay Kit, prepared by mixing the assay buffer and substrate, should be stored at -80°C in the dark and remains effective for at least one year. For short-term storage, it is recommended to store at -20°C in the dark and use within 3-6 months.
Only for research and not intended for treatment of humans or animals
Journals Using SBS Genetech Products Universities Using SBS Genetech Products
SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory
