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  3. D-Luciferin potassium

D-Luciferin potassium  (Synonyms: D-荧光素钾盐; D-(-)-Luciferin potassium; Firefly luciferin potassium; Beetle Luciferin potassium)

目录号: HY-12591B 纯度: 99.97%
COA 产品使用指南

D-Luciferin 是荧光素酶 (Luc) 的天然底物,对萤火虫产生典型的黄绿光进行催化。该反应产生的 560 nm 化学发光在几秒钟内达到峰值,当底物荧光素过量时,光输出与荧光素酶浓度成正比。荧光素酶 (luc) 基因是研究和活性分子筛选的常用报告基因。化学发光技术实际上是无背景的,使得 luc 报告基因成为检测低水平基因表达的理想选择。在标准荧光计数仪中可以可靠地测量到 0.02 pg 的荧光素酶。除了用于基因表达的外,荧光素酶还用于 ATP 的检测。MCE提供萤火虫荧光素酶 (HY-P1004) 、荧光素游离酸 (HY-12591A) 及其水溶性钠盐 (HY-12591) 和钾盐 (HY-12591B) 。

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D-Luciferin potassium Chemical Structure

D-Luciferin potassium Chemical Structure

CAS No. : 115144-35-9

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10 mM * 1 mL in Water ¥550
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5 mg ¥312
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10 mg ¥500
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50 mg ¥900
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100 mg ¥1250
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250 mg ¥1800
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500 mg ¥2500
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1 g ¥3500
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Other Forms of D-Luciferin potassium:

MCE 顾客使用本产品发表的 42 篇科研文献

IF

    D-Luciferin potassium purchased from MCE. 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 potassium purchased from MCE. 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 potassium purchased from MCE. 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 potassium purchased from MCE. 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 potassium purchased from MCE. 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.
    • 生物活性

    • 实验参考方法

    • 纯度 & 产品资料

    • 参考文献

    生物活性

    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 offer 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) .

    体外研究
    (In Vitro)

    1. 操作前注意事项
    a) D-Luciferin 在 100 mM 的水性缓冲液 (pH 6.1-6.5) 中易于溶解。储备溶液可在不含 ATP 的水中制成,并储存在 -20℃ 的温度下,以防光线照射。游离酸必须用适当的碱中和以溶解。在较高的 pH 值下,荧光素经历碱催化的脱氢脲醛生成,以及外消旋成L-异构体。
    b) D-Luciferin 可用于任何现有的报告基因检测或 ATP 检测系统。
    c) 如果检测 ATP,需要戴手套和使用不含 ATP 的容器,尽量减少所有可能的 ATP 污染源。只使用无菌的不含 ATP 的水和试剂。所有试剂制剂使用蒸压水。
    2. 实验操作
    此说明仅提供实验参考,具体实验方法根据您的具体实验进行更改。
    2.1 体外成像实验示例
    a) 在无菌水中制备100 mM (100-200X) 荧光素储备溶液 拌匀。立即使用或一次性等份使用,并在 -20℃ 下储存,避免冻融循环,避免暴露在阳光下。
    b) 在预热过的组织培养液中配制 0.5-1 mM D-Luciferin 工作液
    c) 除去细胞中的培养基。
    d) 在细胞中加入 D-Luciferin 工作液,成像前 37℃ 孵育细胞 5-10 分钟。
    2.2 体内成像实验示例
    a) 在 DPBS 中制备 15 mg/mL 荧光素储备溶液,不含 Mg2+Ca2+ 拌匀。
    b) 过滤器通过 0.2μm 过滤器对溶液进行灭菌。立即使用,或一次性等份使用,并在 -20℃ 下储存,避免冻融循环,避免暴露在阳光下。
    c) 成像前 10-15 分钟,以 150 mg/kg (或 10 μL/g 荧光素储备溶液) 的动物体重腹腔注射 D-Luciferin。
    注:应为每个动物模型进行 D-Luciferin 动力学研究,以确定峰值信号时间。
    2.3 基因检测实验示例
    a) 在无菌水中制备100 mM 荧光素储备溶液。立即使用,或一次性等份使用,并在 -20℃ 下储存,避免冻融循环,避免暴露在阳光下。
    b) 在 25 mM tricine 缓冲液 (pH 7.8) 中制备含有 3 mM ATP、1 mM DTT 和 15 mM MgSO4 的 1 mM D-Luciferin 工作溶液。
    c) 用移液管将 5-10 μL 细胞裂解液移到微孔板中。使用不含裂解物的裂解试剂或缓冲液作为空白对照。
    d) 根据说明,使用荧光素工作溶液为光度计注入底部。
    e) 立即注入 200 μL D-Luciferin工作溶液,结合时间为10秒。

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

    体内研究
    (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.

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

    分子量

    318.41

    Formula

    C11H7KN2O3S2

    CAS 号
    性状

    固体

    颜色

    Light yellow to yellow

    中文名称

    D-萤光素钾盐

    运输条件

    Room temperature in continental US; may vary elsewhere.

    储存方式

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

    *In solvent : -80°C, 2 years; -20°C, 1 year (sealed storage, away from moisture and light)

    溶解性数据
    细胞实验: 

    H2O 中的溶解度 : 17.5 mg/mL (54.96 mM; 超声助溶 (<60°C))

    配制储备液
    浓度 溶剂体积 质量 1 mg 5 mg 10 mg
    1 mM 3.1406 mL 15.7030 mL 31.4060 mL
    5 mM 0.6281 mL 3.1406 mL 6.2812 mL
    查看完整储备液配制表

    * 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效
    储备液的保存方式和期限:-80°C, 2 years; -20°C, 1 year (sealed storage, away from moisture and light)。-80°C储存时,请在2年内使用, -20°C储存时,请在1年内使用。

    * 备注:如您选择水作为储备液,请稀释至工作液后,再用 0.22 μm 的滤膜过滤除菌后使用。

    • 摩尔计算器

    • 稀释计算器

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

    质量
    =
    浓度
    ×
    体积
    ×
    分子量 *

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

    This equation is commonly abbreviated as: C1V1 = C2V2

    浓度 (start)

    C1

    ×
    体积 (start)

    V1

    =
    浓度 (final)

    C2

    ×
    体积 (final)

    V2

    动物实验:

    以下溶解方案,请直接配制工作液。建议现用现配,在短期内尽快用完。 以下溶剂前显示的百分比是指该溶剂在您配制终溶液中的体积占比; 如在配制过程中出现沉淀、析出现象,可以通过加热和/或超声的方式助溶。

    • 方案 一

      请依序添加每种溶剂: PBS

      Solubility: 8.33 mg/mL (26.16 mM); 澄清溶液; 超声助溶 (<60°C)

    动物溶解方案计算器
    请输入动物实验的基本信息:

    给药剂量

    mg/kg

    动物的平均体重

    g

    每只动物的给药体积

    μL

    动物数量

    由于实验过程有损耗,建议您多配一只动物的量
    计算结果
    工作液所需浓度 : mg/mL
    该产品水溶性佳,请具体参考实测 水 / PBS / Saline 中的溶解度数据。
    您所需的储备液浓度超过该产品的实测溶解度,如有需要,请与 MCE 中国技术支持联系。
    纯度 & 产品资料

    纯度: 99.97%

    染色示例
    参考文献
    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. 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.

    参考文献

    完整储备液配制表

    * 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效
    储备液的保存方式和期限:-80°C, 2 years; -20°C, 1 year (sealed storage, away from moisture and light)。-80°C储存时,请在2年内使用, -20°C储存时,请在1年内使用。

    可选溶剂 浓度 溶剂体积 质量 1 mg 5 mg 10 mg 25 mg
    H2O 1 mM 3.1406 mL 15.7030 mL 31.4060 mL 78.5151 mL
    5 mM 0.6281 mL 3.1406 mL 6.2812 mL 15.7030 mL
    10 mM 0.3141 mL 1.5703 mL 3.1406 mL 7.8515 mL
    15 mM 0.2094 mL 1.0469 mL 2.0937 mL 5.2343 mL
    20 mM 0.1570 mL 0.7852 mL 1.5703 mL 3.9258 mL
    25 mM 0.1256 mL 0.6281 mL 1.2562 mL 3.1406 mL
    30 mM 0.1047 mL 0.5234 mL 1.0469 mL 2.6172 mL
    40 mM 0.0785 mL 0.3926 mL 0.7852 mL 1.9629 mL
    50 mM 0.0628 mL 0.3141 mL 0.6281 mL 1.5703 mL

    * 备注:如您选择水作为储备液,请稀释至工作液后,再用 0.22 μm 的滤膜过滤除菌后使用。

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