| Size | Price | Stock | Qty |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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Purity: ≥98%
D-Luciferin (free acid) is a popular and cell-permeable bioluminescent substrate of luciferase in the presence of ATP with a Km of approximately 2 μM, it is used in luciferase-based bioluminescence imaging and cell-based high-throughput screening applications. D-luciferin could emit lights upon oxidative decarboxylation in the presence of ATP. D-luciferin provides a bioluminescent signal for in vivo and in vitro detection of cellular ATP levels. D-Luciferin chould be used to assay the expression of the luciferase gene linked to a promoter of interest. Alternatively, D-luciferin and luciferase can be used to assess ATP availability in cellular or biochemical assays. D-luciferin could be administrated intravenously or intraperitonealy.
| Targets |
Natural substrate of luciferase (Luc) enzyme
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| ln Vitro |
1. Note:
a) D-luciferin exists three forms: free acid, potassium salt, and sodium salt. The sodium and potassium forms of D-luciferin can be easily dissolved in aqueous buffer solutions (pH 6.1-6.5). The stock solution can be prepared with ATP free water and stored in the dark (protect from light) at -20 ° C. While D-luciferin free acid must be neutralized with an appropriate base in order to dissolve. At higher pH values, luciferin will form dehydro-luciferin under alkaline catalysis and racemize into L-luciferin. b) D-fluorescein can be used in any existing report analysis or ATP analysis system. c) If testing ATP, please wear gloves and use ATP free containers to minimize all possible sources of ATP contamination. Only use sterile ATP free water and reagents. Prepare all reagents using high-pressure sterilized water. 2. Experimental protocol: This protocol may be adjusted to meet your specific requirements, as it only serves as a guide. 2.1 Example of In vitro Bioluminescence Image Analysis. a) Prepare D-luciferin stock solution in DMSO. Mix well. Immediately use or aliquot it, store at -20 ℃, avoid repeated freezing and thawing, and avoid exposure to light. b) Prepare 0.5-1 mM D-Luciferin working solution and preheat the tissue culture medium. c) Suck out the culture medium from the cultured cells. d) Add D-luciferin working solution to the cells and incubate them at 37 ℃ for 5-10 minutes before imaging. 2.2 In vivo experiments Due to the large dose used for in vivo experiments, generally 150 mg/kg, it is recommended to choose potassium salt or sodium salt of D-luciferin. 2.3 Example for Fluorescein Reporter Gene Testing a) Prepare D-luciferin stock solution in DMSO. Immediately use or aliquot it, store at -20 ℃, avoid repeated freezing and thawing, and avoid exposure to light. b) Prepare 1 mM D-Luciferin working solution and 3 mM ATP, dissolve 1 mM DTT and 15 mM MgSO4 in 25 mM tricine buffer at pH 7.8. c) Transfer 5-10 μ L of cell lysate to a microplate. Use lysis reagents or buffer solutions without lysate as blank. d) According to the manufacturer's instructions, infuse the luminescence meter with D-Luciferin working solution. e) Immediately inject 200 μ L of D-Luciferin working solution, with an integration time of 10 seconds |
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| ln Vivo |
The most popular method at the moment is bioluminescence imaging (BLI), which uses D-luciferin as a substrate and firefly luciferase (Fluc) as a reporter gene. A time-intensity curve was created by graphing the total signal intensity versus the amount of time following D-luciferin injection. Apart from the peak signal, a surrogate signal for the peak signal was identified at specific time intervals (5, 10, 15, and 20 min) following D-luciferin injection. To depict the pattern of temporal changes following D-luciferin injection, the signal in a given time-intensity curve is normalized against the peak signal in the curve [3]. Intraperitoneal or intravenous injection: Use 10 μL of D-Luciferin stock solution per gram of body weight. An injection of 150 mg/kg should typically contain 200 μL for a 20 g mouse. After thawing at room temperature, dissolve D-Luciferin (sodium or potassium salt) in dPBS (without calcium or magnesium) until the final concentration reaches 15 mg/mL. After passing 5–10 mL of sterile HO through a 0.22 µM filter, discard the water. Pass the D-luciferin solution through a 0.22 µM syringe filter that has been produced.
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| Enzyme Assay |
D-luciferin is the natural substrate of all luciferases that catalyze the production of light in bioluminescent insects. The present review covers the synthesis of D-luciferin and derivatives or analogues that are substrates or inhibitors of the luciferase from the American firefly Photinus pyralis, the enzyme more frequently used in techniques of in vitro and optical imaging[1].
Brain-derived neurotrophic factor (BDNF) plays a crucial role in numerous brain functions, including memory consolidation. Previously, we generated a Bdnf-Luciferase transgenic (Bdnf-Luc) mouse strain to visualize changes in Bdnf expression using in vivo bioluminescence imaging. We successfully visualized activity-dependent Bdnf induction in living mouse brains using a d-luciferin analog, TokeOni, which distributes to the brain and produces near-infrared bioluminescence. In this study, we compared the patterns of bioluminescence signals within the whole body of the Bdnf-Luc mice produced by d-luciferin, TokeOni and seMpai, another d-luciferin analog that produces a near-infrared light. As recently reported, hepatic background signals were observed in wild-type mice when using TokeOni. Bioluminescence signals were strongly observed from the region containing the liver when using d-luciferin and TokeOni. Additionally, we detected signals from the brain when using TokeOni. Compared with d-luciferin and TokeOni, signals were widely detected in the whole body of Bdnf-Luc mice by seMpai. The signals produced by seMpai were strong in the regions containing skeletal muscles in particular. Taken together, the patterns of bioluminescence signals in Bdnf-Luc mice vary when using different luciferase substrates. Therefore, the expression of Bdnf in tissues and organs of interest could be visualized by selecting an appropriate substrate.[4] |
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| Cell Assay |
Organic anion transporter 1 (SLC22A6/OAT1) plays a key role in renal tubular excretion of endo- and exogenous anionic substances including drugs. Since the inhibition of OAT1 function by a concomitant drug may cause pharmacokinetic drug-drug interactions (DDIs) in clinical practice, an in vitro uptake study to evaluate the inhibition potency of OAT1 is useful for the prediction and avoidance of DDIs and recommended for drug candidates in drug development. In this chapter, we describe a rapid and highly sensitive functional assay of OAT1 based on bioluminescence (BL) detection using D-luciferin as a substrate in living cells. The principle of measurement simply relies on the biochemical feature of D-luciferin to be recognized as a substrate of OAT1, and the BL intensity depending on intracellular D-luciferin level and luciferase activity, thereby allowing the quantitative analysis of OAT1-mediated D-luciferin transport. The BL measurement can be completed within 1 min without experimental procedures for removing extracellular uptake solution and washing cells, both of which involve in the conventional uptake studies using isotope-labeled or fluorescent compounds. The present method is applicable to high-throughput screening to identify and avoid potential OAT1 inhibitors in drug development[5].
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| Animal Protocol |
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| References |
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| Additional Infomation |
Photinus luciferin is a 1,3-thiazolemonocarboxylic acid consisting of 3,5-dihydrothiophene-4-carboxylic acid having a 6-hydroxybenzothiazol-2-yl group at the 2-position. It has a role as a luciferin. It is a member of benzothiazoles, a 1,3-thiazolemonocarboxylic acid and an imidothioate. It is a conjugate acid of a Photinus luciferin(1-). It is an enantiomer of an ent-Photinus luciferin.
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| Molecular Formula |
C11H8N2O3S2
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| Molecular Weight |
280.32
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| Exact Mass |
279.997
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| Elemental Analysis |
C, 47.13; H, 2.88; N, 9.99; O, 17.12; S, 22.87
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| CAS # |
2591-17-5
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| Related CAS # |
D-Luciferin sodium;103404-75-7;D-Luciferin potassium;115144-35-9
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| PubChem CID |
92934
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| Appearance |
Typically exists as White to yellow solids at room temperature
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| Density |
1.8±0.1 g/cm3
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| Boiling Point |
587.6±60.0 °C at 760 mmHg
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| Melting Point |
200-204ºC
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| Flash Point |
309.2±32.9 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.865
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| LogP |
0.87
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
18
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| Complexity |
391
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| Defined Atom Stereocenter Count |
1
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| SMILES |
S1C(C2=NC3C([H])=C([H])C(=C([H])C=3S2)O[H])=N[C@@]([H])(C(=O)O[H])C1([H])[H]
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| InChi Key |
BJGNCJDXODQBOB-SSDOTTSWSA-N
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| InChi Code |
InChI=1S/C11H8N2O3S2/c14-5-1-2-6-8(3-5)18-10(12-6)9-13-7(4-17-9)11(15)16/h1-3,7,14H,4H2,(H,15,16)/t7-/m1/s1
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| Chemical Name |
(4S)-2-(6-hydroxy-1,3-benzothiazol-2-yl)-4,5-dihydrothiazole-4-carboxylic acid
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.92 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 2: ≥ 2.08 mg/mL (7.42 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (7.42 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5674 mL | 17.8368 mL | 35.6735 mL | |
| 5 mM | 0.7135 mL | 3.5674 mL | 7.1347 mL | |
| 10 mM | 0.3567 mL | 1.7837 mL | 3.5674 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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