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

1. Prepare samples (50 µL)
2. Add 50 µL Gaussia Luciferase working solution
3. Incubate at room temperature for 10 - 15 minutes
4. Monitor luminescence intensity

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

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

Transfer 50 µL (for Cat#12530), 0.5 mL (for Cat#12531) or 2.5 mL (for Cat#12532) of Reaction Buffer (Component B) into 1 vial of Luciferase Substrate (Component A), and mix well. Note: Prior to addition, centrifuge briefly before opening the vial of Luciferase Substrate (Component A).

Dilute 100X Gaussia Luciferase assay stock solution using 1:100 dilution factor with Assay Buffer (Component C) to prepare Gaussia Luciferase working solution.
Note        The reconstituted Gaussia luciferase working solution is very sensitive to light. It is not stable, should be prepared fresh, kept on ice, and used within 2 hours.

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. For blank wells (medium without the cells), add the corresponding amount of compound buffer.

2. Incubate the cell plates in a 37 °C, 5% CO₂ incubator for a desired period of time, typically 4 hours to overnight.

3. Run Gaussia Luciferase Assay: Pipette 50 µL/well/96-well plate or 12.5 µL/well/384-well plate of the serial diluted Gaussia Luciferase or culture supernatant into a microtiter plate, and then mix with 50 µL/well/96-well plate or 12.5 µL/well/384-well plate of the Gaussia Luciferase working solution.
4. Incubate the plate at room temperature for 10 to 15 minutes, protected from light.
5. Read luminescence intensity with a luminometer.