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  4. D-Luciferin sodium

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

Cat. No.: HY-12591 Purity: 99.92%
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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 sodium Chemical Structure

D-Luciferin sodium Chemical Structure

CAS No. : 103404-75-7

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Based on 41 publication(s) in Google Scholar

Other Forms of D-Luciferin sodium:

D-Luciferin sodium Related Antibodies

Top Publications Citing Use of Products

36 Publications Citing Use of MCE D-Luciferin sodium

IF

    D-Luciferin sodium 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 sodium 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 sodium 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 sodium 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 sodium 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) The D-luciferin salts are readily soluble in aqueous buffers (pH 6.1-6.5) up to 100 mM. Stock solutions can be made in ATP-free water and stored at -20°C, protect from light. The free acid must be neutralized with an appropriate base to solubilize. 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.
    The following protocol is an example for potassium and sodium salt preparation. It can be adapted for most cell types and in vivo animal use.
    2.1 Example protocol for in vitro bioluminescent image assays
    a) Prepare a 100 mM (100-200X) Luciferin stock solution in sterile water. Mix well. Use immediately, or make single use aliquots, and store at -20°C, 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 Luciferin working solution to cells, and incubate the cells for 5-10 minutes at 37°C just prior to imaging.
    2.2 Example protocol for in vivo bioluminescent image assays
    a) Prepare a 15 mg/mL Luciferin stock solution in DPBS, without Mg2+ and Ca2+. Mix well.
    b) Filter sterilizes the solution through a 0.2 μm filter. Use immediately, or make single use aliquots, and store at -20°C, avoid freeze-thaw cycles, avoid exposure to the light.
    c) Inject the luciferin intra-peritoneally (i.p.) 10-15 minutes before imaging at 150 mg/kg (or 10 μL/g of luciferin stock solution) of the animal body weight.
    Note: A kinetic study of luciferin should be performed for each animal model to determine peak signal time.
    2.3 Example protocol for luciferin reporter assays
    a) Prepare a 100 mM Luciferin stock solution in sterile water. Use immediately, or make single use aliquots, and store at -20°C, 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 luciferin working solution according to manufacturer’s instructions.
    e) Inject 200 μL of 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.

    D-Luciferin sodium Related Antibodies
    In Vivo

    Bioluminescence imaging (BLI) using the firefly luciferase (Fluc) as a reporter gene and D-luciferin as a substrate is currently the most widely employed technique. The total signal intensity is plotted against the time after D-luciferin injection to generate a time-intensity curve. In addition to the peak signal, the signals at fixed time points (5, 10, 15, and 20 min) after D-luciferin injection are determined as alternatives to the peak signal. The signal in a given time-intensity curve is normalized for the peak signal in the curve to represent the pattern of temporal changes after D-luciferin injection[3].
    Inject with 10 μL of D-luciferin (intraperitoneally or intravenously) stock solution per gram of body weight: normally ~200 μL for a 20 g mouse for a standard 150 mg/kg injection.
    Thaw D-Luciferin (either Potassium or Sodium Salt) at room temperature and dissolve in dPBS (no calcium or magnesium) to a final concentration of 15 mg/mL. Pre-wet a 0.22 μm filter by drawing through 5-10 mL of sterile H2O and discard water. Sterilize the D-Luciferin solution through the prepared 0.22 μm syringe filter.

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

    302.30

    Formula

    C11H7N2NaO3S2
    CAS No.
    Appearance

    Solid

    SMILES

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

    Room temperature in continental US; may vary elsewhere.
    Storage

    4°C, sealed storage, away from moisture and light

    *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

    Solvent & Solubility
    In Vitro: 

    H2O : 250 mg/mL (826.99 mM; ultrasonic and warming and heat to 60°C)

    DMSO : 100 mg/mL (330.80 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 3.3080 mL 16.5399 mL 33.0797 mL
    5 mM 0.6616 mL 3.3080 mL 6.6159 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 (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    * Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

    • 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    40% PEG300    5% Tween-80    45% Saline

      Solubility: ≥ 2.5 mg/mL (8.27 mM); Clear solution

      This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 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.
    • Protocol 2

      Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

      Solubility: ≥ 2.5 mg/mL (8.27 mM); Clear solution

      This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

      Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.

    For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
    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:  PBS

      Solubility: 100 mg/mL (330.80 mM); Clear solution; Need ultrasonic

    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.
    Calculation results:
    Working solution concentration: mg/mL
    This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.
    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).
    Purity & Documentation

    Purity: 99.92%

    COA (246 KB) HNMR (199 KB) LCMS (111 KB)

    Handling Instructions (2659 KB)
    Dyeing Example
    References
    Animal Administration
    [3]

    Mice[3]
    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 (sealed storage, away from moisture and 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
    DMSO / H2O 1 mM 3.3080 mL 16.5399 mL 33.0797 mL 82.6993 mL
    5 mM 0.6616 mL 3.3080 mL 6.6159 mL 16.5399 mL
    10 mM 0.3308 mL 1.6540 mL 3.3080 mL 8.2699 mL
    15 mM 0.2205 mL 1.1027 mL 2.2053 mL 5.5133 mL
    20 mM 0.1654 mL 0.8270 mL 1.6540 mL 4.1350 mL
    25 mM 0.1323 mL 0.6616 mL 1.3232 mL 3.3080 mL
    30 mM 0.1103 mL 0.5513 mL 1.1027 mL 2.7566 mL
    40 mM 0.0827 mL 0.4135 mL 0.8270 mL 2.0675 mL
    50 mM 0.0662 mL 0.3308 mL 0.6616 mL 1.6540 mL
    60 mM 0.0551 mL 0.2757 mL 0.5513 mL 1.3783 mL
    80 mM 0.0413 mL 0.2067 mL 0.4135 mL 1.0337 mL
    100 mM 0.0331 mL 0.1654 mL 0.3308 mL 0.8270 mL

    * Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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    D-Luciferin sodium Related Classifications

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    • Do most proteins show cross-species activity?

      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.

    Keywords:

    D-Luciferin103404-75-7D-(-)-LuciferinFirefly luciferinBeetle LuciferinFluorescent DyeInhibitorinhibitorinhibit

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