Laurdan
(Synonyms: 6-十二酰基-N,N-二甲基-2-萘胺) 目录号 : GC18338Laurdan是一种极性敏感脂膜荧光探针，用于对细胞膜中的脂筏（也称为脂质微域）进行成像，Laurdan的吸收波长为340-380nm，发射波长为440-490nm。
Cas No.:74515-25-6
Sample solution is provided at 25 µL, 10mM.
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Related Biological Data
Cell membrane homeostasis analysis of Z. bailii. (D) Membrane fluidity.
Laurdan (GLPBIO, China) was added to the suspension at a final concentration of 1 μM and incubated for 60 min at 37 °C away from light.
J Agr Food Chem (2023). PMID: 37929595 IF: 6.1004 -
Related Biological Data
Continued Cinacalcet (CNA) kills bacteria by disrupting membrane and energy metabolism and by producing ROS.(f and g) Characterization of membrane fluidity by laurdan staining.
MRSA with different treatments was analysed for membrane permeability using SYTOX Green, membrane fluidity using laurdan (GlpBio) and membrane potential using DiSC3(5).
J Antimicrob Chemoth (2024): dkae051. PMID: 36016574 IF: 5.6
Quality Control & SDS
- View current batch:
- Purity: >98.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
This plan only provides a guide, please modify it to meet your specific needs.
1. Solution preparation
(1) Storage solution: Dissolve Laurdan in DMSO or chloroform to a final concentration of 5 mM.
Note: After unused storage solution is aliquoted, store it at -20℃ in the dark to avoid repeated freezing and thawing.
(2) Working solution: Dilute the storage solution with experimental buffer (e.g. cell culture medium) to the required working concentration, usually in the range of 1-10 µM.
Note: Please adjust the optimal working concentration according to the actual situation or refer to the literature to set the gradient concentration by yourself. The working solution must be prepared and used immediately.
2. Steps for measuring the generalized polarization (GP) of Bacillus subtilis cell membrane by Laurdan [1] (from the literature, for reference only)
(1) Cell treatment: Cultivate cells to the middle of the logarithmic growth phase, wash them four times in PBS containing 2% glucose and 1% DMF, resuspend them in the same buffer, and adjust the OD600 to 0.3.
(2) Liposome preparation: Bacillus subtilis polar lipid extract was prepared by detergent dialysis, and the prepared liposome solution was squeezed through a 0.4μm filter membrane 20 times. The 10mg/ml liposome stock solution was diluted to a concentration of 1 mg/ml with Tris buffer (5mM, pH 7.4).
(3) Liposome labeling: Add Laurdan solution to the cell suspension to a final concentration of 10 μM and incubate in the dark for 30 minutes.
(4) Fluorescence measurement: Laurdan fluorescence measurement was performed using a BioTek Synergy MX plate reader. The excitation wavelength was 350 nm, the emission wavelengths were 460 nm and 500 nm, and readings were taken every 2 minutes.
(5) Laurdan fluorescence polarization calculation: Laurdan generalized polarization (GP) value was calculated using the following formula: (I460-I500)/(I460+I500).
Note: This experimental protocol only provides a reference for a Laurdan generalized polarization (GP) measurement experiment of cell membrane. Please modify the working concentration and staining time according to experimental needs or refer to the literature.
References:
[1]Saeloh D, Tipmanee V, Jim K K, et al. The novel antibiotic rhodomyrtone traps membrane proteins in vesicles with increased fluidity[J]. PLoS pathogens, 2018, 14(2): e1006876.
Laurdan is a polarity-sensitive lipid membrane fluorescent probe used to image lipid rafts (also known as lipid microdomains) in cell membranes, the absorption wavelength of Laurdan is 340-380nm and the emission wavelength is 440-490nm[1, 2, 3]. Laurdan is membrane-embedded. Its hydrophobic tail can be inserted into the phospholipid bilayer of the cell membrane, while the polar head remains near the membrane surface, which can detect polarity changes and the presence of water molecules inside the membrane [4]. Laurdan is phase-sensitive. Its fluorescence emission in the liquid crystal phase will show a large red shift, while it will show a small red shift in the gel phase, which can detect the phase state and fluidity of the membrane [5]. Laurdan can measure the generalized polarization (GP) of the cell membrane. High GP is usually associated with low fluidity, low polarity or high cholesterol content of the membrane, while low GP is the opposite [6].
References:
[1] Horváth Á, Erostyák J, Szőke É. Effect of Lipid Raft Disruptors on Cell Membrane Fluidity Studied by Fluorescence Spectroscopy[J]. International Journal of Molecular Sciences, 2022, 23(22): 13729.
[2] Alvares S M, Dunn C A, Brown T A, et al. The role of membrane microdomains in transmembrane signaling through the epithelial glycoprotein Gp140/CDCP1[J]. Biochimica et Biophysica Acta (BBA)-General Subjects, 2008, 1780(3): 486-496.
[3] Goldys E M. Fluorescence applications in biotechnology and life sciences[M]. John Wiley & Sons, 2009.
[4] Jay A G, Hamilton J A. Disorder amidst membrane order: standardizing laurdan generalized polarization and membrane fluidity terms[J]. Journal of fluorescence, 2017, 27: 243-249.
[5] Bianchetti G, Azoulay-Ginsburg S, Keshet-Levy N Y, et al. Investigation of the membrane fluidity regulation of fatty acid intracellular distribution by fluorescence lifetime imaging of novel polarity sensitive fluorescent derivatives[J]. International Journal of Molecular Sciences, 2021, 22(6): 3106.
[6] Yu W, So P T, French T, et al. Fluorescence generalized polarization of cell membranes: a two-photon scanning microscopy approach[J]. Biophysical journal, 1996, 70(2): 626-636.
Laurdan是一种极性敏感脂膜荧光探针，用于对细胞膜中的脂筏（也称为脂质微域）进行成像，Laurdan的吸收波长为340-380nm，发射波长为440-490nm[1, 2, 3]。Laurdan具有膜嵌入性，它的疏水尾部可以插入细胞膜的磷脂双层中，而极性头部留在膜表面附近，能够探测膜内部的极性变化和水分子存在情况[4]。Laurdan具有相态敏感性，在液晶相中的荧光发射会显示较大的红移，而在凝胶相中则会显示较小的红移，能够检测膜的相态和流动性[5]。Laurdan可测量细胞膜的广义极化（GP），高GP通常与膜的低流动性、低极性或高胆固醇含量相关，而低GP则相反[6]。
| Cas No. | 74515-25-6 | SDF | |
| 别名 | 6-十二酰基-N,N-二甲基-2-萘胺 | ||
| 化学名 | 1-[6-(dimethylamino)-2-naphthalenyl]-1-dodecanone | ||
| Canonical SMILES | CN(C)C1=CC=C2C(C=CC(C(CCCCCCCCCCC)=O)=C2)=C1 | ||
| 分子式 | C24H35NO | 分子量 | 353.5 |
| 溶解度 | Chloroform: 10 mg/ml | 储存条件 | Store at -20°C,protect from light |
| General tips | 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 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.8289 mL | 14.1443 mL | 28.2885 mL |
| 5 mM | 0.5658 mL | 2.8289 mL | 5.6577 mL |
| 10 mM | 0.2829 mL | 1.4144 mL | 2.8289 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。