Double-Luciferase Reporter Assay Kit
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Price:
US$797.50
(Size: 50 rxns)
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Double-Luciferase Reporter Assay Kit provides an efficient means of detecting the activities of firefly (photinus pyralis) and renilla (sea pansy) luciferases, using luciferin and coelenterazine as substrates respectively.
Kit contents: | Component | 50 rxns | 200 rxns |
| Luciferase Reaction Buffer 1 | 5 ml | 20 ml |
| Luciferase Reaction Buffer 2 | 5 ml | 20 ml |
| Luciferase Reaction Substrate 1 | 1 vial | 4 vials |
| Luciferase Reaction Substrate 2 (50X) | 100 µl | 400 µl |
| Cell Lysis Buffer (5X) | 5 ml | 20 ml |
Reagents required but not provided: - PBS (1X)
- Nuclease-Free Water
| Target |
Double-Luciferase Reporter Assay Kit |
| Storage |
Store at -20 °C. Once prepared (see Assay Procedure), store the Luciferase Reaction Reagents 1 and 2 in the dark at -20 °C for up to one month or -70 °C for up to 1 year. |
| Availability |
Shipped within 10-20 working days. |
| Note |
This product is for research use only. This product is shipped with dry ice. |
| Directions for use |
Assay Procedure - Bring the Luciferase Reaction Buffer 1 and Luciferase Reaction Buffer 2 to room temperature. If precipitates are observed in Luciferase Reacton Buffer 2, mix well and/or heat in a 37 °C water bath until they are dissolved.
- Prepare the Luciferase Reaction Reagent 1 by adding 5 ml of Luciferase Reaction Buffer 1 into 1 vial of Luciferase Reaction Substrate 1. Mix well and store in the dark at -20 °C for up to 1 month, or at -70 °C for up to 1 year. Avoid repeated freeze/thaw cycles.
- Prepare the Luciferase Reaction Reagent 2 by adding Luciferase Reaction Substrate 2 to Luciferase Reaction Buffer 2 at a ratio of 1:49 (e.g. add 50 µl of Luciferase Reaction Substrate 2 to 2.45 ml of Luciferase Reaction Buffer 2). Mix well and store in the dark at -20 °C for up to 1 month, or at -70 °C for up to 1 year. Avoid repeated freeze/thaw cycles.
- Prepare the 1X Cell Lysis Buffer by adding 5X Cell Lysis Buffer to Nuclease-Free Water at a ratio of 1:4 (e.g. add 1 ml of 5X Cell Lysis Buffer to 4 ml of Nuclease-Free Water).
- Remove the culture medium. Carefully rinse twice with 1X PBS and add an appropriate volume of 1x Cell Lysis Buffer according to the table below. Fully lyse cells at room temperature for 10 minutes. Scrape the cells into a microcentrifuge tube, then centrifuge at 12,000 × g for 10 minutes. Take the supernatant, which contains the cell lysate.
| Cell Culture Plate Type | Lysis Buffer per Well | | 6-well | 500 µl | | 12-well | 250 µl | | 24-well | 100 µl | | 48-well | 60 µl | | 96-well | 20 µl | - Bring the Luciferase Reaction Reagent 1 and Luciferase Reaction Reagent 2 to room temperature.
- Add 100 µl of Luciferase Reaction Reagent 1 (equilibrated to room temperature) into a microcentrifuge tube or well in an opaque 96-well plate. Add 20 µl of cell lysate and mix thoroughly. Measure the luminescence using a luminometer to measure the firefly luciferase activity.
- Add 100 µl of Luciferase Reaction Reagent 2 (equilibrated to room temperature) into a microcentrifuge tube or well in an opaque 96-well plate. Add 20 µl of cell lysate and mix thoroughly. Measure the luminescence using a luminometer to measure the renilla luciferase activity.
Notes: - It is recommended to add samples and reagents using a multichannel pipette in steps 7 and 8, when measuring a large number of samples.
- Luciferase Reaction Reagents 1 and 2 are easily oxidized. It is recommended to use Luciferase Reaction Reagents 1 and 2 immediately after they are equilibrated to room temperature.
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Research Articles on Double-Luciferase Reporter Assay Kit
(1 Publications and Reviews)
