Firefly luciferase is a protein with a molecular weight of about 61 kD that catalyzes the oxidation of the substrate luciferin to oxyluciferin in the presence of ATP, oxygen and magnesium ions. During the oxidation of Luciferin, a bioluminescence with a wavelength of about 560 nm is emitted, which can be determined by a luminometer or liquid flash analyzer. With luciferin and luciferase as bioluminescent systems, gene expression can be detected very sensitively and efficiently. Generally, the transcriptional regulatory element of the gene of interest or the 5' promoter region is cloned upstream of luciferase, or the 3'-UTR region is cloned downstream of luciferase, etc., to construct a reporter gene plasmid, transfect the cells and treat the cells with appropriate drugs, and finally determine the luciferase activity of the cell lysate and judge the transcriptional regulation effect of drug treatment on the target gene by the level of luciferase activity (Fig 1).
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The Single Firefly Luciferase Reporter Gene Assay Kit is a highly sensitive, highly signal-stable one-step assay for direct chemiluminescence determination of intracellular Firefly luciferase activity without washing or collecting cells.
The firefly luciferase detection reagent provided in this kit can be added to the cell culture plate in the same volume as the culture medium for 5 minutes to react, and chemiluminescence detection can be performed. This kit is flexible and convenient to use, has high detection sensitivity, stable luminescence signal, wide linear range of measured samples, and better performance than major domestic similar products (Fig 2). In addition, this product can also be used for the detection of lysing and collecting preserved cell samples.
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