KeyTec® Firefly-Renilla Luciferase Detection Kit

KeyTec® Firefly-Renilla luciferase Detection kit is designed for the sensitive and robust detection of Firefly-Renilla dual luciferase reporter gene assays. The Firefly reagent detects Firefly luciferase activities, enabling accurate measurement of genetic element regulation. Additionally, the Renilla reagent quenches the Firefly luminescent signal and detects Renilla luciferase activities for normalization. This normalization can control for specific or nonspecific cellular responses and transfection efficiencies. The detection principle is based on Luminescent technology. Within the kit, D-Luciferin reacts with Firefly luciferase, generating a sensitive luminescence signal with a half-life of approximately 2 hours. Addition of the KeyTec® Renilla Luciferase Stop & reaction buffer quenches the Firefly reaction's luminescence, and simultaneously, Coelenterazine reacts with Renilla luciferase, generating a luminescence signal that can also be read within 2 hours. The process provides a highly sensitive, robust, and homogeneous assay for the detection of Firefly-Renilla luciferase reporter gene expression in mammalian cells.

Use KeyTec® Firefly-Renilla Luciferase Detection Kit to detect the overexpression of Firefly luciferase and Renilla luciferase in the HEK293 cell line. The results showed a linear relationship between the luminescence signal and the number of cells. HEK293 cells, cultured in DMEM medium with 10% FBS, were serially diluted two-fold, starting from 50,000 cells per well in a 96-well plate. Perform the assay according to the procedure outlined in “Assay Procedure”. Ten minutes after adding the Firefly luciferase detection reagent, measure the luminescence signal using the Envision's Luminescence program. (Example program details include Mirror: Luminescence, Em filter: Luminescence 700, Measurement height: 6.5 mm, and Measurement time: 1 s). Then ten minutes after adding the Renilla luciferase detection reagent, measure the luminescence signal using the Envision's Luminescence program.