Nano-Lux Luciferase Assay System

The Nano-Lux Luciferase Assay Substrate is mixed with the Nano-Lux Luciferase Assay Buffer at a 1:50 volume ratio to prepare the Detection Reagent. The prepared reagent is then added directly to the cell culture at a volume equal to the sample volume. It lyses the cells, releases intracellular Nano-Lux luciferase and initiates the luminescent reaction. Following a 3–5-minute incubation at room temperature, luminescence is measured using a microplate reader equipped with a luminescence detection module. The generated luminescent signal has a half-life of up to 2 hours under suitable assay conditions, providing flexibility when measuring multiple samples or plates. As the Detection Reagent is added directly to the cell culture, there is no requirement to remove the culture medium or wash the cells before detection.

The Nano-Lux Luciferase Assay System can be used for:

• Gene regulation
• Signal transduction
• Protein stability
• Viral infection and replication
• Promoter and pathway activity

• High Sensitivity: Optimized chemistry strengthens Nano-Lux luminescence while reducing background signals.
• Wide Detection Window: Supports the measurement of both low and high reporter expression levels.
• Stable Light Output: Provides a luminescent signal with a half-life of up to two hours.
• Simple Homogeneous Workflow: Add the reagent directly to cultured cells, incubate for 3–5 minutes, and measure luminescence.
• No Cell Washing Required: Cells can be cultured and assayed in the same multiwell plate.
• Suitable for High-Throughput Assays: Compatible with opaque multiwell plates and automated liquid-handling workflows.

Storage Conditions: Store all components at −30°C to −15°C. Transport at 0°C or below.

Working Reagents: Prepare Detection Reagent immediately before use by combining Substrate and Buffer at 1:50 volume ratio. Prepare only the amount required for the experiment.

Reagent Preparation:

1. Thaw Nano-Lux Luciferase Assay Buffer in room-temperature water bath not exceeding 25°C, or overnight at 2–8°C. Allow Buffer to equilibrate fully to room temperature. Substrate remains liquid and does not require thawing.
2. Briefly centrifuge Nano-Lux Luciferase Assay Substrate vial before opening to collect contents at bottom.
3. Prepare Detection Reagent by combining Substrate and Buffer at 1:50 volume ratio. For example: add 200 µL Substrate to 10 mL Buffer. Mix gently and thoroughly. Avoid vigorous shaking.

Detection Steps:

1. Remove cell-culture plate from incubator and allow it to equilibrate at room temperature for approximately 10 minutes.
2. Add a volume of Nano-Lux Luciferase Detection Reagent equal to the volume of test sample already present in each well (e.g., add 100 µL reagent to 100 µL cell culture).
3. Allow plate to stand at room temperature for 3–5 minutes. During this incubation, the Detection Reagent lyses cells, releases intracellular Nano-Lux luciferase and initiates luminescent reaction.
4. Measure luminescent signal using microplate reader equipped with luminescence detection module. For optimal sensitivity, use open or full-spectrum luminescence detection. When defined wavelength is required, detect at approximately 460 nm.