Description
D-Luciferin Sodium Salt is a highly soluble, bioluminescent substrate for firefly luciferase (FLuc)-based assays in both in vitro and in vivo applications.
Many researchers prefer the sodium salt form for its rapid dissolution and excellent aqueous solubility, simplifying preparation and ensuring consistent dosing, especially in longitudinal or high-throughput imaging studies.
With extensive validation in published papers, it remains a dependable tool for tracking gene expression, tumor progression, and cellular behavior in real time. Supported by decades of bioluminescence research and GoldBio’s commitment to value and quality, it’s the ideal choice for laboratories seeking precision, reliability, and cost-effective performance.
Firefly Luciferin is identical to Beetle Luciferin. (See "Additional Information" for structures.)
Proven and Published is a Registered trademarks in the U.S. Patent and Trademark Office.
Functional Highlights and Mechanism
-
The substrate (D-Luciferin) is oxidized by firefly luciferase in the presence of ATP, Mg²⁺ and O₂, resulting in emission of visible light. This underlies luminescence-based quantification of reporter gene activity or cell viability.
-
The sodium salt form ensures high solubility in aqueous buffers, facilitating preparation of stock solutions for cell-based assays or in vivo injections with minimal solubility issues.
-
Because light emission is directly proportional to luciferase activity, which in turn depends on ATP availability. This reagent serves as a sensitive indicator of cellular viability, gene expression, or signaling activity in luciferase-based systems.
-
In live-animal imaging, the substrate enables non-invasive monitoring of luciferase-expressing cells (tumor, stem cell, infectious models) over time, supporting longitudinal studies of disease progression or drug efficacy.
-
The “Proven and Published®” label indicates that this reagent has been used in peer-reviewed work and comes with supporting documentation (handbooks, protocols) from GoldBio, which aids reproducibility and user confidence.
Recommended Applications and Usage Notes
Recommended Applications
-
Bioluminescence imaging (BLI) of luciferase-expressing cells or animal models (e.g., mice or rats).
-
In vitro luciferase reporter assays used in gene expression studies, promoter activity analysis, signal transduction monitoring or high-throughput screening.
-
ATP- or energy-dependent cell viability or cytotoxicity assays using luciferase systems.
-
Kinetic or endpoint assays where light output is the read-out for enzyme activity, reporter gene expression or metabolic status.
-
Longitudinal in vivo monitoring in preclinical models (e.g., tumor growth, stem cell engraftment, infectious disease) where reproducible luciferin substrate performance matters.
Usage Tips and Considerations
-
Prepare a sterile, high-concentration stock solution (for example 15 mg/mL in DPBS or sterile buffer) and filter sterilize (0.22 µm) if used for in vivo injections. GoldBio’s “Luciferin In Vivo Handbook” recommends such procedures.
-
For in vivo work, optimize dose (commonly ~150 mg/kg for mice, but adjust based on model) and timing (there is a delay between injection and maximal luminescence) for your specific system.
-
When using in vitro assays, ensure that your buffer contains sufficient ATP and Mg²⁺, as luciferase reaction requires these cofactors. GoldBio’s recommended assay buffer (TMCA) includes 100 mM Tris-HCl (pH 7.8), 5 mM MgCl₂, 0.25 mM CoA (hydrate), 0.15 mM ATP.
-
Protect the reagent and prepared stocks from light and avoid repeated freeze/thaw cycles to maintain activity and reproducibility.
-
Because light output and kinetics depend on temperature, pH, substrate concentration and presence of other components, run pilot experiments to fine-tune your protocol and verify linearity of signal in your system.
-
Confirm the lot-specific Certificate of Analysis from GoldBio to verify purity, stability, and suitability for your application.
Common Applications
(Click each for more information)
Substrate for Firefly Luciferase in Bioluminescent Imaging
-
Purpose: To visualize gene expression and biological processes in living organisms using light-emitting luciferase systems.
-
How It Works: D-Luciferin is oxidized by firefly luciferase in an ATP-dependent reaction, producing oxyluciferin, AMP, CO₂, and visible light (~560 nm). The sodium salt provides high aqueous solubility, facilitating in vivo use.
-
Applications: In vivo imaging of tumors, gene therapy vectors, and transgenic models.
Fraga, H. (2008). Firefly luminescence: A historical perspective and recent developments. Photochemical & Photobiological Sciences, 7(2), 146–158.
Monitoring Tumor Progression in Oncology Models
-
Purpose: To track the growth and spread of luciferase-tagged cancer cells over time without sacrificing animals.
-
How It Works: Luciferase-expressing tumor cells emit bioluminescence when exposed to D-Luciferin, allowing real-time, noninvasive monitoring of tumor burden using CCD-based imaging. This approach enables longitudinal tracking of tumor progression and therapeutic response.
-
Applications: Xenograft studies, metastasis tracking, therapeutic efficacy evaluation.
Edinger, M., et al. (1999). Noninvasive assessment of tumor cell proliferation in animal models by bioluminescence imaging. Cancer Research, 59(23), 5982–5986.
Tracking Cell Fate and Engraftment in Regenerative Medicine
-
Purpose: To follow the localization, survival, and function of luciferase-expressing cells after transplantation.
-
How It Works: Upon systemic or localized administration of D-Luciferin, only viable cells expressing luciferase catalyze light production, which can be imaged in real time.
-
Applications: Stem cell therapy studies, CAR-T cell tracking, and tissue engineering research.
Cao, F., et al. (2006). In vivo visualization of embryonic stem cell survival, proliferation, and migration after cardiac delivery. Circulation, 113(7), 1005–1014. https://doi.org/10.1161/CIRCULATIONAHA.105.576025
Quantifying Promoter Activity and Gene Regulation
-
Purpose: To measure transcriptional activity by linking gene promoters to luciferase reporters.
-
How It Works: The amount of bioluminescent signal emitted upon D-Luciferin oxidation correlates with the activity of the promoter upstream of the luciferase gene.
-
Applications: Gene regulation studies, drug screening, pathway activation profiling.
Brasier, A. R., et al. (1998). Luciferase reporter gene assay: quantitative analysis of gene expression. Methods in Molecular Biology, 102, 259–273. https://doi.org/10.1385/0-89603-413-5:259
High-Throughput Screening in Drug Discovery
-
Purpose: To detect cellular responses, enzyme activity, or pathway inhibition in large-scale screens.
-
How It Works: Luciferase-based biosensors emit a quantifiable light signal in response to chemical or biological stimuli when D-Luciferin is present.
-
Applications: Cell-based screening assays for inhibitors, activators, or toxic compounds.
Fan, F., & Wood, K. V. (2007). Bioluminescent assays for high-throughput screening. Assay and Drug Development Technologies, 5(1), 127–136. https://doi.org/10.1089/adt.2006.053
Application Summary
-
in vivo cellular imaging
- luciferase and ATP assays
- gene reporter assays
- high throughput sequencing
- contamination assays
Storage/Handling
Store desiccated at -20°C. Protect from light.
GoldBio Luciferase Assay Buffer
| TMCA Buffer |
Final Conc. |
| Tris-HCl, pH 7.8 |
100mM
|
| MgCl2
|
5mM |
| Coenzyme A (CoA) (hydrate) |
0.25mM |
| ATP (disodium salt hydrate) |
0.15mM |
For the full recipe and instructions, go to GoldBio's Luciferin in vitro Handbook.
Powered by Bioz
