Description
GoldBio’s D-Luciferin Potassium Salt is a high-purity, water-soluble substrate used for bioluminescent imaging of firefly luciferase activity in live cells and animal models. With proven performance in published studies, this reagent is essential for tracking gene expression, tumor progression, stem cell migration, and real-time cellular events.
The potassium salt form ensures excellent solubility and bioavailability, making it ideal for in vivo imaging and high-throughput luciferase assays. Whether you’re conducting non-invasive optical imaging or measuring promoter activity in vitro, GoldBio’s D-Luciferin Potassium Salt delivers bright, consistent signal and unmatched reliability across bioluminescent applications.
Tested and Certified for in vivo imaging (See "Luciferin FAQ" in additional information)
Firefly Luciferin is identical to Beetle Luciferin. (See "Additional Information" for structures.)
"LUCK" and "Proven and Published" are Registered trademarks in the U.S. Patent and Trademark Office.
Functional Highlights and Mechanism
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D-Luciferin is the natural substrate of firefly luciferase (FLuc). In the presence of ATP, Mg²⁺, and oxygen, luciferase catalyzes luciferin adenylation followed by oxidative decarboxylation, emitting visible light (λₘₐₓ ≈ 560 nm in vitro, shifting toward red in vivo due to tissue absorption).
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Because the potassium salt form dissolves readily, it allows for accurate preparation of stock solutions, which supports reproducible luminescent profiling in cell culture or live-animal imaging.
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The luminescent output is proportional to luciferase activity, making luciferin a powerful reporter reagent for gene expression, promoter activity, cell viability (via luciferase reporters), and in vivo tracking of luciferase-expressing cells or pathogens.
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Because bioluminescence signals are inherently low-background (no external excitation source as in fluorescence), using a high-quality luciferin substrate enhances the signal-to-noise ratio and supports sensitive experiments such as small-tumor detection or stem-cell tracking.
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GoldBio supports the reagent with detailed handbooks for in-vitro and in-vivo use (preparation, injection, kinetics), enabling users to implement robust workflows.
Recommended Applications and Usage Notes
Recommended Applications
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In vivo bioluminescent imaging (BLI) of luciferase-expressing animals (e.g., mouse models of cancer, infectious disease, stem-cell engraftment).
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In vitro luciferase reporter gene assays (cell culture, multi-well plates) to monitor promoter activity, gene regulation, signal transduction, or compound screening.
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High-throughput screening (HTS) assays that rely on luciferase output as a read-out for functional genomics or drug discovery.
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Monitoring cell viability, cytotoxicity or metabolic state via luciferase reporters in cells or small animal systems.
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Longitudinal studies where non-invasive imaging of the same animals or samples over time is needed, supported by the stable and well-characterized luciferin reagent.
Usage Tips and Considerations
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Prepare a fresh luciferin stock solution by dissolving the potassium salt in sterile, oxygen-equilibrated buffer or saline (e.g., 15 mg/mL in DPBS) and filter sterilize (0.22 µm) if used for in vivo injection.
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For animal imaging, an intraperitoneal injection of ~150 mg/kg (depending on model) is common; optimize dose, injection route, and timing to determine peak luminescence for your system.
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Because the luciferase-luciferin reaction depends on ATP, Mg²⁺ and oxygen, ensure your system has sufficient cofactors and is well-oxygenated for optimal light output.
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For cell-based or plate-based assays, minimize light leakage, use dark adaptation if needed, and keep exposure times consistent for comparison across treatments.
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Store the product desiccated at -20 °C and protect from light; avoid repeated freeze-thaw of stock solutions to maintain consistency.
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When using in vivo models, consider pharmacokinetics and biodistribution of luciferin — timing between injection and imaging may vary by species or tissue.
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Always refer to the lot-specific Certificate of Analysis supplied by GoldBio to ensure reagent performance and purity for your experiments.
Common Research Applications
(Click each for more information)
Substrate for Firefly Luciferase in Bioluminescent Imaging
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Purpose: To enable ATP-dependent light production for monitoring gene expression, cell proliferation, and molecular interactions.
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How It Works: D-Luciferin serves as the natural substrate for firefly luciferase (FLuc). In the presence of ATP, Mg²⁺, and O₂, luciferase catalyzes the adenylation and oxidation of luciferin to form oxyluciferin, AMP, PPi, and CO₂, emitting visible light (λₘₐₓ ≈ 560 nm).
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Applications: In vivo imaging of tumors, transgene expression, cell trafficking in small animals.
Bhaumik, S., & Gambhir, S. S. (2002). Optical imaging of Renilla luciferase reporter gene expression in living mice. Proceedings of the National Academy of Sciences, 99(1), 377–382.
Non-Invasive Monitoring of Tumor Progression and Metastasis
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Purpose: To track tumor burden over time in live animal models without sacrificing subjects.
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How It Works: Luciferase-expressing tumor cells emit bioluminescence after administration of D-Luciferin, allowing temporal monitoring of tumor growth and metastasis.
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Applications: Oncology research, therapeutic efficacy evaluation, cancer metastasis models.
Noninvasive assessment of tumor cell proliferation in animal models by bioluminescence imaging.
In Vivo Tracking of Stem Cells and Immune Cells
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Purpose: To visualize the biodistribution and persistence of luciferase-labeled cells after transplantation or injection.
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How It Works: D-Luciferin crosses cell membranes and is oxidized only in viable luciferase-expressing cells, emitting light that can be detected via CCD imaging.
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Applications: Regenerative medicine, CAR-T cell tracking, cell-based therapy studies.
Cao, F., Lin, S., Xie, X., Ray, P., Patel, M., Zhang, X., ... & Wu, J. C. (2006). In vivo visualization of embryonic stem cell survival, proliferation, and migration after cardiac delivery. Circulation, 113(7), 1005–1014.
Promoter Activity and Gene Regulation Studies
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Purpose: To quantify transcriptional activity under various experimental conditions.
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How It Works: Luciferase gene is placed downstream of regulatory sequences; light emission from D-Luciferin oxidation reflects promoter strength or response to transcription factors.
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Applications: Promoter screening, pathway activation analysis, high-throughput drug screening.
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
Bioluminescent High-Throughput Screening (HTS) in Drug Discovery
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Purpose: To enable rapid, sensitive detection of molecular interactions, enzyme activities, or pathway modulation.
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How It Works: Luciferase-coupled reporter systems produce a quantifiable light signal in response to test compounds.
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Applications: GPCR and kinase assays, pathway inhibition screens, cytotoxicity tests.
Fan, F., & Wood, K. V. (2007). Bioluminescent assays for high-throughput screening. Assay and Drug Development Technologies, 5(1), 127–136.
Application Summary
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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 Concentration |
| Tris-HCl, pH 7.8 |
100mM
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| 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.
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