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| Product Name | Dual One-Step Luciferase Reporter Gene Assay Kit (Glow Type) |
| Catalog No. | RESK-0150 |
| Description | This is a glow-type dual luciferase reporter gene assay kit featuring high sensitivity and a stable luminescent signal. The kit contains high-purity firefly luciferin and coelenterazine. In a single sample, firefly luciferase is first detected with firefly luciferin as the substrate; the firefly signal is then quenched while Renilla luciferase is detected with coelenterazine as the substrate. Renilla luciferase serves as an internal control, eliminating variations caused by differences in cell number and transfection efficiency between wells and improving the accuracy of results. Compared with flash-type dual luciferase assays, this kit offers: 1) a more stable luminescent signal, providing greater flexibility in experimental design; 2) better suitability for high-throughput detection without reliance on automatic injectors and with no need to remove culture medium or centrifuge, greatly simplifying the workflow; 3) no obvious pungent odor. The kit is compatible with a variety of common cell culture media, including RPMI 1640, DMEM, MEM-alpha, F12, and DMEM/F12, with a signal half-life of approximately 2 h (at 22 °C) in these media, meeting the needs of most high-throughput experiments. |
| Intended Use | Glow-type dual luciferase reporter gene detection with stable, long-half-life signals, suitable for high-throughput analysis of firefly (primary reporter) and Renilla (internal control) luciferase activities in cultured cells. |
| Principle / Technology | Firefly luciferase (approximately 61 kDa) catalyzes the oxidation of luciferin to oxyluciferin in the presence of ATP, magnesium ions, and oxygen, emitting bioluminescence at approximately 560 nm. Renilla luciferase (approximately 36 kDa) catalyzes the oxidation of coelenterazine to coelenteramide in the presence of oxygen, emitting bioluminescence at approximately 480 nm. Both signals are measured with a luminometer. The kit sequentially detects firefly luciferase first, then quenches the firefly signal while detecting Renilla luciferase, with glow-type kinetics providing a stable, long-lived signal. |
| Detection Method | Glow-type bioluminescence detection using a luminometer or luminescence-capable microplate reader; no automatic injector required. |
| Sample Type | Cultured cells in common culture media (RPMI 1640, DMEM, MEM-alpha, F12, DMEM/F12). Tobacco leaf samples can also be assayed after thorough grinding in liquid nitrogen. |
| Performance Range / Specifications | Signal half-life of approximately 2 h at 22 °C in common culture media. Normal luminescence in common media including DMEM, F12, MEM-alpha, and RPMI-1640, with minimal signal differences between media. Effective quenching of the firefly signal upon addition of the Renilla substrate. |
| Sensitivity / LOD | High sensitivity for both firefly and Renilla luciferase detection. |
| Reaction Conditions / Protocol | Simple add-and-read workflow: add the detection reagent to the sample and read the luminescence. No medium removal, centrifugation, or automatic injector is required. Firefly luciferase is detected first, followed by quenching of the firefly signal and detection of Renilla luciferase in the same sample. |
| Components / Formulation | High-purity firefly luciferin and coelenterazine substrates with corresponding reaction buffers and firefly signal quenching component. |
| Storage Conditions | Store at -25 to -15 °C. Store substrates protected from light at low temperature; coelenterazine is recommended to be stored at -80 °C. |
| Shelf Life | 1 year when stored at -25 to -15 °C. |
| Package Specifications | 100 T 1000 T |
| Molecular Weight | Firefly luciferase: approximately 61 kDa. Renilla luciferase: approximately 36 kDa. |
| Shipping Conditions | Shipped on dry ice. |
| Regulatory / Compliance | For research use only. Not for use in diagnostic procedures. |
| Compatibility | Compatible with common cell culture media including RPMI 1640, DMEM, MEM-alpha, F12, and DMEM/F12; suitable for high-throughput detection formats. |
| Application Notes / Precautions | The two substrates do not require strict light protection during handling, but should be stored protected from light at low temperature; coelenterazine is recommended to be stored at -80 °C. For co-transfection of dual reporter vectors, optimize the ratio by pilot experiments (e.g., firefly to Renilla vector ratios of 1:10, 1:20, 1:50, 1:100); a ratio yielding firefly luminescence higher than Renilla luminescence is preferred. Firefly luciferase serves as the primary reporter and Renilla luciferase as the internal control reporter. For tobacco leaf samples, grind the leaves thoroughly in liquid nitrogen before detection. |
For research use only, not for clinical use.
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