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D-Luciferin  (Synonyms: D-(-)-Luciferin; Firefly luciferin; Beetle Luciferin)

Cat. No.: HY-12591A Purity: 99.54%
COA Handling Instructions

D-luciferin is the natural substrate of the enzyme luciferase (Luc) that catalyzes the production of the typical yellowgreen light of fireflies. The 560 nm chemiluminescence from this reaction peaks within seconds, with light output that is proportional to luciferase concentration when the substrate luciferin is present in excess. The luciferase (luc) gene is a popular reporter gene for research and agent screening. Chemiluminescent techniques are virtually background-free, making the luc reporter gene ideal for detecting low-level gene expression. As little as 0.02 pg of luciferase can be reliably measured in a standard scintillation counter. In addition to its role as a reporter of gene expression, luciferase is commonly used in an extremely sensitive assay for ATP. We of er the firefly luciferase (HY-P1004), luciferin free acid (HY-12591A), as well as its water-soluble sodium salts (HY-12591) and potassium salts (HY-12591B) .

For research use only. We do not sell to patients.

D-Luciferin Chemical Structure

D-Luciferin Chemical Structure

CAS No. : 2591-17-5

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Customer Review

Based on 41 publication(s) in Google Scholar

Other Forms of D-Luciferin:

Top Publications Citing Use of Products

36 Publications Citing Use of MCE D-Luciferin

IF

    D-Luciferin purchased from MedChemExpress. Usage Cited in: Acta Pharm Sin B. 12 March 2022.

    Tumor growth is monitored by bioluminescence imaging. Prior to imaging, D-luciferin substrate is injected intraperitoneally into the mice (150 mg/kg per mouse).

    D-Luciferin purchased from MedChemExpress. Usage Cited in: New Phytol. 2022 Sep 2.  [Abstract]

    ERF16-nLUC and CaM2-cLUC vectors are cotransfected in N. benthamiana leaves. 10 min after the abaxial sides of leaves are sprayed with 1 mM Luciferin.

    D-Luciferin purchased from MedChemExpress. Usage Cited in: Pharmacol Res. 2021 Mar 2;105527.  [Abstract]

    After cell inoculation for 7 days, mice are injected intraperitoneally with D-luciferin substrate at 150 mg/kg and anesthetized 10% chloral hydrate (5 mL/kg).

    D-Luciferin purchased from MedChemExpress. Usage Cited in: Cell Mol Gastroenterol Hepatol. 2021;12(3):839-856.

    HEK293T cells are seeded into a 6-well plate and transfected with in vitro synthesized mRNA containing the luciferase gene by Lipofectamine 2000. Luciferase substrate D-Luciferin sodium salt (1 mM) is added into the culture medium immediately after transfection. Luciferase activity is detected on the GloMax 96 Microplate Luminometer.

    D-Luciferin purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2020 Sep 17;11(9):765.  [Abstract]

    At Day 10, 25, and 31, mice are anesthetized with isoflurane and then injected with 75 mg/kg D-Luciferin solution for imaging.
    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

    • Customer Review

    Description

    D-luciferin is the natural substrate of the enzyme luciferase (Luc) that catalyzes the production of the typical yellowgreen light of fireflies. The 560 nm chemiluminescence from this reaction peaks within seconds, with light output that is proportional to luciferase concentration when the substrate luciferin is present in excess. The luciferase (luc) gene is a popular reporter gene for research and agent screening. Chemiluminescent techniques are virtually background-free, making the luc reporter gene ideal for detecting low-level gene expression. As little as 0.02 pg of luciferase can be reliably measured in a standard scintillation counter. In addition to its role as a reporter of gene expression, luciferase is commonly used in an extremely sensitive assay for ATP[1]. We of er the firefly luciferase (HY-P1004), luciferin free acid (HY-12591A), as well as its water-soluble sodium salts (HY-12591) and potassium salts (HY-12591B) .

    In Vitro

    1. Precautions
    a) There are three forms of D-luciferin, namely free acid (HY-12591A), potassium salt (HY-12591B) and sodium salt (HY-12591). The sodium salt and potassium salt forms of D-luciferin are easily soluble in aqueous buffers (pH 6.1-6.5). Stock solutions can be made in ATP-free water and stored at -20℃, protect from light. The free acid can be neutralized with DMSO or an appropriate base to dissolve. At a higher pH, luciferin undergoes a base-catalyzed formation of dehydroluciferin, as well as racemization to the L-isomer.
    b) The D-luciferin can be used with any existing reporter assay or ATP assay system.
    c) If testing for ATP, minimize all possible sources of ATP contamination by wearing gloves and using ATP-free containers. Use only sterile ATP-free water and reagents. Use autoclaved water for all reagent preparations.
    2. Experimental Protocols
    This protocol only provides a guideline, and should be modified according to your specific needs.
    2.1 Example protocol for in vitro bioluminescent image assays
    a) Prepare D-luciferin stock solution in DMSO. Mix well. Use immediately, or make single use aliquots, and store at -20℃, avoid freeze-thaw cycles, avoid exposure to the light.
    b) Prepare a 0.5-1 mM working solution of D-Luciferin in pre-warmed tissue culture medium.
    c) Aspirate media from cultured cells.
    d) Add D-Luciferin working solution to cells, and incubate the cells for 5-10 minutes at 37℃ just prior to imaging.
    2.2 In vivo assays
    Note: Since the in vivo experimental dose is relatively large, generally 150 mg/kg, it is recommended to choose HY-12591B or HY-12591.
    2.3 Example protocol for D-Luciferin reporter assays
    a) Prepare D-luciferin stock solution in DMSO. Use immediately, or make single use aliquots, and store at -20℃, avoid freeze-thaw cycles, avoid exposure to the light.
    b) Prepare a 1 mM working solution of D-Luciferin with 3 mM ATP, 1 mM DTT and 15 mM MgSO4 in 25 mM tricine buffer pH 7.8.
    c) Pipette 5-10 μLof cell lysate into a microplate. Use lysis reagent or buffer without lysate as a blank.
    d) Prime luminometer with D-Luciferin working solution according to manufacturer’s instructions.
    e) Inject 200 μL of D-luciferin working solution with no delay and a 10 second integration time.

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

    Molecular Weight

    280.32

    Formula

    C11H8N2O3S2

    CAS No.
    Appearance

    Solid

    Color

    White to yellow

    SMILES

    O=C([C@@H]1N=C(C2=NC3=CC=C(O)C=C3S2)SC1)O

    Structure Classification
    Initial Source

    insect

    Shipping

    Room temperature in continental US; may vary elsewhere.

    Storage

    4°C, protect from light

    *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

    Solvent & Solubility
    In Vitro: 

    DMSO : 125 mg/mL (445.92 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    H2O : 9.09 mg/mL (32.43 mM; ultrasonic and adjust pH to 9 with 1M NaOH)

    H2O : < 0.1 mg/mL (insoluble)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 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
    View the Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    • Molarity Calculator

    • Dilution Calculator

    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    Mass
    =
    Concentration
    ×
    Volume
    ×
    Molecular Weight *

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

    ×
    Volume (start)

    V1

    =
    Concentration (final)

    C2

    ×
    Volume (final)

    V2

    In Vivo:

    Select the appropriate dissolution method based on your experimental animal and administration route.

    For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
    To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
    The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

    • Protocol 1

      Add each solvent one by one:  10% DMSO    90% Corn Oil

      Solubility: ≥ 2.5 mg/mL (8.92 mM); Clear solution

      This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

    • Protocol 2

      Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

      Solubility: ≥ 2.08 mg/mL (7.42 mM); Clear solution

      This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

      Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

    Animal weight
    (per animal)

    g

    Dosing volume
    (per animal)

    μL

    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Please enter your animal formula composition:
    %
    DMSO +
    +
    %
    Tween-80 +
    %
    Saline
    Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
    The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
    Calculation results:
    Working solution concentration: mg/mL
    Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

    *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

    The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
    Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
     If the continuous dosing period exceeds half a month, please choose this protocol carefully.
    Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
    Purity & Documentation

    Purity: 99.87%

    Dyeing Example
    References
    Animal Administration
    [2]

    Mice[2]
    In vivo BLI is performed using a cooled charge-coupled device camera system (IVIS Imaging System 100) 3, 5, 7, 10, 12, 14, 19, 21, 24, and 28 days after the inoculation of HCT116-Luc cells. Mice are injected with 75 mg/kg D-luciferin in 100 μL of phosphate-buffered saline subcutaneously near the scapula and were placed in the light-tight chamber of the imaging system. Beginning 5 min after injection, dorsal luminescent images with an exposure time of 1 s are acquired sequentially at a rate of one image per min until 20 min after D-luciferin injection. Data acquisition is continued until 40 min postinjection on days 3 or 5 and until 25 min on day 7, because of the prolonged time course of light emission. Binning is 4 and the field of view is 15 cm.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    References

    Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    H2O / DMSO 1 mM 3.5674 mL 17.8368 mL 35.6735 mL 89.1838 mL
    5 mM 0.7135 mL 3.5674 mL 7.1347 mL 17.8368 mL
    10 mM 0.3567 mL 1.7837 mL 3.5674 mL 8.9184 mL
    15 mM 0.2378 mL 1.1891 mL 2.3782 mL 5.9456 mL
    20 mM 0.1784 mL 0.8918 mL 1.7837 mL 4.4592 mL
    25 mM 0.1427 mL 0.7135 mL 1.4269 mL 3.5674 mL
    30 mM 0.1189 mL 0.5946 mL 1.1891 mL 2.9728 mL
    DMSO 40 mM 0.0892 mL 0.4459 mL 0.8918 mL 2.2296 mL
    50 mM 0.0713 mL 0.3567 mL 0.7135 mL 1.7837 mL
    60 mM 0.0595 mL 0.2973 mL 0.5946 mL 1.4864 mL
    80 mM 0.0446 mL 0.2230 mL 0.4459 mL 1.1148 mL
    100 mM 0.0357 mL 0.1784 mL 0.3567 mL 0.8918 mL
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      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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