Product Description
Firefly luciferase is a protein with a molecular weight of approximately 61 kD that can catalyze the oxidation of Luciferin to Oxyluciferin in the presence of ATP, magnesium ions, and oxygen. During the oxidation of Luciferin, bioluminescence is emitted. Sea kidney luciferase is a protein with a molecular weight of approximately 36 kD that can catalyze the oxidation of Coelenterazine to Coelenteramide in the presence of oxygen. During the oxidation of Coelenterazine, it also emits biological fluorescence. Biological fluorescence can be measured using a chemiluminescence analyzer (Luminometer) or a liquid scintillation analyzer.
In DLR detection, the activities of firefly luciferase and Renilla luciferase can be sequentially detected in a single sample. Firstly, Luciferin was used as the substrate to detect Firefly Luciferase, followed by Coelenterazine as the substrate to detect Renilla Luciferase. Subsequently, when adding the substrate for Renilla Luciferase, substances that inhibit the catalytic luminescence of Firefly Luciferase were also added, so that only the activity of Renilla Luciferase could be detected in subsequent tests, achieving dual luciferase reporter gene detection. Through the bioluminescence system of luciferase and its substrates, gene expression can be detected very sensitively and efficiently. Usually, the transcription regulatory elements or 5 'promoter regions of the genes of interest are cloned upstream of Luciferase, or the 3' - UTR region is cloned downstream of Luciferase to construct a reporter gene plasmid. Then, cells are transfected, treated with appropriate drugs, and lysed to determine luciferase activity. Judging the transcriptional regulation of target genes by drug treatment and other factors based on the level of luciferase activity. Renilla Luciferase is relatively more commonly used as an internal reference for transfection efficiency to eliminate differences in cell number and transfection efficiency.
The Firefly&Renilla dual luciferase reporter gene detection kit has the characteristics of rapid detection, high sensitivity, wide detection range, and no interference from endogenous cellular activity.
Product components
Kit components | 100T | 1000T |
A.5× Passive Luciferase Lysis Buffer | 10 mL | 100 mL |
B.Firefly Luciferase Assay Buffer | 10 mL | 100 mL |
C.D-Luciferin | 2 mg | 20 mg |
D.Renilla Luciferase Assay Buffer | 10 mL | 100 mL |
E.50× Coelenterazine | 200 µL | 2 mL |
Transportation and storage
Blue ice transportation- Stored at 20 ℃, with a shelf life of 12 months.
[Note]: It is recommended to use the B group distribution system as a 2 mg/mL storage solution for component C. Component B, component D, and component C prepared as a storage solution should be divided into small batches according to experimental needs. All testing working fluids are recommended to be prepared and used immediately to avoid repeated freezing and thawing.
This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.
Back
