Amplite® Renilla Luciferase Reporter Gene Assay Kit *Maximized Luminescence*

Protocol summary

1. Prepare cell plates
2. Treat cells as needed
3. Remove medium from cell plates
4. Add Renilla Luciferase working solution (100 µL/well for 96-well plate or 25 µL/well for 384-well plate)
5. Incubate at room temperature for 5 - 10 minutes
6. Monitor luminescence intensity

Important notes
Thaw all the kit components to room temperature before use. For all luminescent experiments, it is recommended to use white plates to get the best results.

1. Add one volume of 100X Luciferase Substrate (Component A) to 100 volumes of Assay Buffer (Component B) to make Renilla Luciferase working solution. Note: The reconstituted Renilla Luciferase working solution is very sensitive to light, should be kept from light. In addition, it is not stable, should be prepared fresh, kept on ice and used within 2 hours.

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

1. Treat cells (or samples) with test compounds by adding 10 µL of 10X test compounds (96-well plate) or 5 µL of 5X test compounds (384-well plate) in desired compound buffer.
    
2. Incubate the cell plate in a 5% CO₂ incubator at 37°C for desired period of time, typically 4 hours to overnight.
   
   
3. Remove the medium completely.
   
   
4. Add 100 µL (96-well plate) or 25 µL (384-well plate) per well of Renilla Luciferase working solution.
   
   
5. Incubate the plate at room temperature for 5 - 10 minutes. Protect from light.
   
   
6. Monitor luminescence intensity with a luminometer.