Description
Duo-Lux Luciferase Assay System (ASRV00017)
Duo-Lux Luciferase Assay System is a next-generation, high-sensitivity, and homogeneous reagent solution engineered for the rapid, concurrent quantitation of two reporter enzymes, Firefly luciferase and Renilla luciferase, from a single cell sample. Designed for simplicity, the convenient, two-step "Add-Mix-Read" protocol generates both stable luminescent signals directly from cells cultured in medium, eliminating the need for pre-conditioning, washing, or centrifugation steps.
| Product Name: | Duo-Lux Luciferase Assay System |
| SKU: | ASRV00017 |
| Size: | 10ml, 100ml |
| Detection Principle: | Luminescence |
| Step | Protocol |
| 1 | Remove the cell culture plate to be tested from the incubator and equilibrate at room temperature for 30 min. |
| 2 |
Firefly (Primary Reporter) Detection: Add an equal volume of Duo-Lux Luciferase Detection Reagent to the cell culture medium to be tested and then mix well. a. Example 96-well: Add 75ul reagent to 75ul cell culture medium b. Example 384-well: Add 20ul reagent to 20ul cell culture medium |
| 3 | Measure: Incubate at RT for up to 10 minutes and detect the Firefly luminescence signal. |
| 4 |
Renilla Detection: Add an equal volume of Duo-Lux Stop & Lux Detection Reagent to the well, based on the original culture volume, and mix it well. a. Example 96-well: Add 75ul reagent to 75ul cell culture medium b. Example 384-well: Add 20ul reagent to 20ul cell culture medium |
| 5 | Measure: Incubate at RT for up to 10 minutes and detect the Renilla luminescence signal. |
|
Detection Step Note 1: The Duo-Lux Stop & Lux reagent must be added to the well within 4 hours after the Duo-Lux Luciferase reagent was added (Step 2). Detection Step Note 2: The Renilla luminescence must be measured in the same plate order as that of the Firefly signal to maintain data integrity. |
|
| Step | Protocol |
| Thawing of Duo-Lux Buffer | Incubate the Duo-Lux buffer at 2 to 8℃ or room temperature to thaw. The product can also be incubated at 22℃ in a water bath but the temperature should not exceed 25℃. |
| Preparation of Duo-Lux Luciferase Detection Reagent | Add the entire bottle of thawed Duo-Lux Luciferase Buffer into the Duo-Lux Luciferase Substrate. Gently invert and mix it well for 3 to 5 times until the substrate is dissolved thoroughly. |
| Preparation of Duo-Lux Stop & Lux Detection Reagent |
Calculate the volume of reagent required for the experiment. Dilute the Duo-Lux Stop & Lux Substrate into the corresponding volume of Duo-Lux Stop & Lux Buffer at a ratio of 1:100. Gently invert and mix the solution well. a. Example: To prepare 5 ml of Duo-Lux Stop & Lux detection reagent, add 50ul of Duo-Lux Stop & Lux Substrate into 5 ml of Duo-Lux Stop & Lux Buffer. |
| Temperature Equilibration | Before use, ensure that both the Duo-Lux Luciferase Buffer and Duo-Lux Stop & Lux Buffer have been equilibrated to room temperature. • Frozen Reagents: If the working Duo-Lux Detection Reagents were stored at -20℃ or -70℃, they should be gently inverted and mixed well 3-5 times after thawing and before use. |
| Product | Code | Contents |
| Duo-Lux Luciferase Assay System (10ml) | ASRV00017-10 |
Duo-Lux Luciferase Buffer 10ml Duo-Lite Luciferase Substrate (lyophilized) 1 vial Duo-Lux Stop & Lux Buffer 10ml Duo-Lux Stop & Lux Substrate 100ul |
| Duo-Lux Luciferase Assay System (100ml) | ASRV00017-100 |
Duo-Lux Luciferase Buffer 100ml Duo-Lite Luciferase Substrate (lyophilized) 1 vial Duo-Lux Stop & Lux Buffer 100ml Duo-Lux Stop & Lux Substrate 1ml |
