Bio FIREFLYGLO luciferase reporter gene detection kit

Cat. No.: G1375755

GRADE & PURITY BioReagent, for Chemiluminescence

Overview

Firefly Luciferase Reporter Gene Assay Kit (Glow) is a glow-type quantitative detection kit characterized by high sensitivity and stable luminescent signals. It is suitable for high-throughput detection of firefly luciferase expression in mammalian cells. This product significantly enhances luminescent signal intensity, effectively improving detection sensitivity. It can be used to detect firefly luciferase reporter gene expression in ADCC Reporter Bioassays or cells.

G1375755	Component	10 mL	100 mL	Storage
G1375755A	Firefly Luciferase Reaction Buffer (Glow)	10 mL	100 mL	-20℃. Store in the dark.
G1375755B	Firefly Luciferase Substrate (Glow)	1 vial	1 vial	-20℃. Store in the dark.

​he main features and differences among three One-Step Firefly Luciferase Reporter Gene Assay Kits from Aladdin are as follows:

For high luminescent signal requirements and detection within 30 minutes: Recommended:  Firefly Luciferase Reporter Gene Assay Kit (Glow Bright) (Cat. No. B1375760)

For both luminescent signal and stability requirements, suitable for batch or continuous operation: Recommended: Firefly Luciferase Reporter Gene Assay Kit  (Cat. No. F773892)

For standard luminescent signal requirements: Recommended:  Firefly Luciferase Reporter Gene Assay Kit (Glow) (Cat. No. G1375755)

For routine laboratory detection, the  Firefly Luciferase Reporter Gene Assay Kit (Glow Bright) (Cat. No. B1375760) is the recommended priority.

Product Cat. No.	Product Name	Luminescence Type	Signal Intensity	Sensitivity	Half-Life	Packaging	Suitable for Batch Samples	Optimal Use After Mixing & Storing at -20°C
F773892	Firefly Luciferase Reporter Gene Assay Kit	Glow	Relatively High	Relatively Sensitive	Up to 2h	1 bottle Buffer 1 vial Substrate	Very Suitable	1 Month
G1375755	Firefly Luciferase Reporter Gene Assay Kit (Glow)	Glow	High	Sensitive	25-30min	1 bottle Buffer 1 vial Substrate	Very Suitable	1 Week
B1375760	Firefly Luciferase Reporter Gene Assay Kit (Glow Bright)	Glow	Very High	Highly Sensitive	25-30min	1 bottle Buffer 1 vial Substrate	Very Suitable	1 Week

Instructions for Use:

1. User-Supplied Materials

• PBS

• Multichannel pipette

• White or black opaque cell culture microplate

• Luminometer or microplate reader equipped with a luminescence detection module.

2. Preparation Before Assay
(1) Initial Reconstitution: Upon first use, transfer the entire contents of the Firefly Luciferase Reaction Buffer (Glow) bottle into the Firefly Luciferase Substrate (Glow) vial. Mix thoroughly until the substrate is completely dissolved. Aliquot the reconstituted detection reagent as needed. For long-term storage, aliquot and store at -70°C. For short-term storage, store aliquots at -20°C for no longer than one month. Use reconstituted reagent promptly.
(2) Pre-assay: Before each experiment, equilibrate the aliquoted, frozen detection reagent to room temperature.

3. Procedure
(1) Equilibrate Plates: Remove the cell culture plate from the incubator and allow it to stand at room temperature for 5-15 minutes.
Note: Use white or black opaque cell culture plates to minimize signal interference between wells.
(2) Add Detection Reagent: Using a multichannel pipette, add the equilibrated Firefly Luciferase Reaction Buffer (Glow) containing the substrate to each cell culture well. The volume added should be equal to the volume of the culture medium in the well. For example:
* 96-well plate: Typically add 80-100 µL culture medium; add 80-100 µL detection reagent.
* 384-well plate: Typically add 20-30 µL culture medium; add 20-30 µL detection reagent.
(3) Mix to Lyse Cells: To ensure complete cell lysis, place the plate on an orbital shaker or an instrument with shaking capability. Shake at medium to high speed at room temperature for 5 minutes.
Note: A shaking time of 5 minutes is recommended. This can be adjusted based on cell density to ensure complete lysis and stable luminescent results.
(4) Detection: Following the shaking step, immediately measure firefly luciferase reporter gene activity using a luminometer or microplate reader.
Note: For optimal results, perform the detection as soon as possible after adding the detection reagent.

Precautions

1. Instrument Selection: Any instrument capable of detecting chemiluminescence is suitable for use with this kit. However, background signal levels and measured values for identical samples may vary between different detectors. Values obtained from different instruments for the same sample are not directly comparable. To prevent well-to-well interference, the use of opaque white or black cell culture plates is strongly recommended.

2. Consistent Conditions: Luminescent signals can be influenced by environmental factors such as medium components and temperature. Ensure consistent detection conditions across all samples within an experiment.

3. Temperature Sensitivity: The enzymatic reaction is temperature-sensitive. It is essential to equilibrate both the detection reagent and the cell culture plate to room temperature before adding the reagent and performing the assay.

4. Reagent Stability: To maintain the stability of the luciferase assay reagent, aliquot it appropriately after reconstitution. For long-term storage, store aliquots at -70°C protected from light. For short-term storage, store at -20°C for no longer than one month. Use aliquots promptly and avoid repeated freeze-thaw cycles.

5. Multiple Plate Assay: If detecting multiple cell culture plates simultaneously, strive to maintain consistent incubation time after adding the detection reagent to each plate before reading. This ensures optimal and comparable results across plates.

6. Emission Wavelength: The bioluminescence catalyzed by firefly luciferase exhibits a peak emission wavelength of 560 nm.

Storage and Shipping

Storage

Protected from light,Store at -20°C

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20°C long term (12 months). Store in the dark. After reconstitution, the Firefly Luciferase Substrate (Glow) can be stored at -70℃ for 12 months, or at -20℃ for up to 1 month. Store in the dark.

Images

Firefly Luciferase Reporter Gene Assay : Firefly Luciferase Reporter Gene Assay Kit (Glow) (G1375755)

Firefly Luciferase Reporter Gene Assay Kit (Glow) (G1375755) -Performance on serially diluted HEK293 cells (50 to 200,000)

The detection performance of this product on serially diluted HEK293 cells stably transfected with the firefly luciferase gene. The data shown in the figure were generated from a 100 μL cell culture sample mixed with 100 μL of detection reagent, followed by incubation on a shaker for 5 minutes. Actual measured data may vary depending on the detection instrument and other factors. Additionally, the maximum detectable cell number can vary considerably with cell type and the intrinsic level of luciferase expression.