Lucifer Yellow CH
(Synonyms: 萤光黄二锂盐,Lucifer Yellow carbohydrazide) 目录号 : GC44089Lucifer Yellow CH是一种水溶性的高强度荧光染料，最大激发/发射分别为430nm/540nm。
Cas No.:67769-47-5
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >80.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
本方案仅提供一个指导，应根据您的具体需要进行修改。
1. 染色液的配制
(1) 制备储存液：采用1mL ddH2O溶解1mg Lucifer Yellow CH配制为1mg/mL的储存液。
注意:未使用的储存液分装后在-20℃或-80°C避光保存，避免反复冻融。
(2) 配制工作液：使用合适的缓冲液（如预热好的无血清细胞培养基）稀释储存液，配制成0.5-1%的Lucifer Yellow CH工作液。
注意:请根据实际情况调整工作液浓度，现用现配。
2. 细胞染色悬浮细胞：离心收集细胞，弃去上清液，用PBS清洗两次，每次5分钟。
贴壁细胞：使用PBS清洗两次，加入胰酶消化细胞，消化完成后经1000g离心3-5分钟。
(1) 在收集好的细胞中加入1mL Lucifer Yellow CH工作液，室温孵育5-30分钟。
(2) 400g，4°C离心3-4分钟，弃去上清。
(3) 使用PBS清洗两次，每次5分钟。
(4) 用1mL无血清培养基或PBS重悬细胞后，在荧光显微镜或流式细胞仪下检测。
注意事项：
1. 荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光淬灭
2. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
Lucifer Yellow CH is a water-soluble, high-intensity fluorescent dye with maximum excitation/emission at 430nm/540nm, respectively. Lucifer Yellow CH contains a carbodihydrazide (CH) group that can covalently bind to surrounding biomolecules during aldehyde fixation. Lucifer Yellow CH is impermeable to cell membranes at physiological pH and can be loaded into living cells by microinjection, pinocytosis, scraping, or ATP-induced permeabilization. Lucifer Yellow CH can be used as a biological tracer to label neural cells, monitor neuronal branching, regeneration, gap junction detection and characterization, and selective ablation of cells [1-5].
References:
[1] Stewart WW. Functional connections between cells as revealed by dye-coupling with a highly fluorescent naphthalimide tracer. Cell. 1978 Jul;14(3):741-59.
[2] Klein M, et al. Transport of lucifer yellow CH into plant vacuoles--evidence for direct energization of a sulphonatedsubstance and implications for the design of new molecular probes. FEBS Lett. 1997 Dec 22;420(1):86-92.
[3] Takeuchi K, et al. Effect of superoxide derived from lucifer yellow CH on voltage-gated currents of mouse taste budcells. Chem Senses. 2008 Jun;33(5):425-32.
[4] Sun Y, Leong N T, Jiang T, et al. Switch-like Arp2/3 activation upon WASP and WIP recruitment to an apparent threshold level by multivalent linker proteins in vivo[J]. Elife, 2017, 6: e29140.
[5] Zhang Y, Hou M C, Li J J, et al. Cardiac β‐adrenergic receptor activation mediates distinct and cell type‐dependent changes in the expression and distribution of connexin 43[J]. Journal of Cellular and Molecular Medicine, 2020, 24(15): 8505-8517.
Lucifer Yellow CH是一种水溶性的高强度荧光染料，最大激发/发射分别为430nm/540nm。Lucifer Yellow CH包含一个碳二酰肼（CH）基团，能够在醛固定过程中与周围的生物分子共价结合。Lucifer Yellow CH在生理pH值下不能渗透细胞膜，可通过显微注射、胞饮作用、刮涂或ATP诱导的渗透作用加载到活细胞中。Lucifer Yellow CH可用作生物示踪剂，标记神经细胞，监测神经元分支、再生、间隙连接检测和表征以及细胞的选择性消融[1-5]。
| Cas No. | 67769-47-5 | SDF | |
| 别名 | 萤光黄二锂盐,Lucifer Yellow carbohydrazide | ||
| 化学名 | 6-amino-2-[(hydrazinylcarbonyl)amino]-2,3-dihydro-1,3-dioxo-1H-benz[de]isoquinoline-5,8-disulfonic acid, dilithium salt | ||
| Canonical SMILES | O=C(C1=CC(S([O-])(=O)=O)=C(N)C2=C1C3=CC(S([O-])(=O)=O)=C2)N(NC(NN)=O)C3=O.[Li+].[Li+] | ||
| 分子式 | C13H9N5O9S2•2Li | 分子量 | 457.2 |
| 溶解度 | 1mg/mL in water | 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 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 2.1872 mL | 10.9361 mL | 21.8723 mL |
| 5 mM | 0.4374 mL | 2.1872 mL | 4.3745 mL |
| 10 mM | 0.2187 mL | 1.0936 mL | 2.1872 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。
Intracellular marking with Lucifer Yellow CH and horseradish peroxidase of cells electrophysiologically characterized as glia in the cerebral cortex of the cat
J Comp Neurol 1979 Jul 15;186(2):173-88.PMID:87405DOI:10.1002/cne.901860205.
Intracellular microelectrodes filled with either Lucifer Yellow CH, a highly florescent dye, or horseradish peroxidase (HRP) were used to electrophysiologically characterize and mark cells in the cerebral cortex of cat. Fifty-eight cells, characterized electrophysiologically as glia, were marked with Lucifer Yellow CH. All were identified as protoplasmic astrocytes, and included cells in the glia limitans of the molecular layer. An additional 54 cells, similarly characterized as glia, were labeled with HRP. The results were the same; only protoplasmic astrocytes were labeled. The "staining quality" of the glia labeled with HRP was superior to that of cells injected with Lucifer Yellow; greater lengths of individual processes were revealed, and they could often be followed to blood vessels where they ended on the walls of vessels with expanded perivascular end-feet. The observations indicate that the many previously reported studies on presumed glial cells in the cat cerebral cortex have characterized the behavior of protoplasmic astrocytes. Neurons were also marked during these experiments. The "staining" quality of the Lucifer Yellow filled neurons was excellent; dendritic spines, axons, and axon collaterals were clearly visible. These fine neuronal details were not as well revealed after HRP labeling. High resting membrane potentials (RMP's) were not a prerequisite for obtaining well-marked neurons (mean RMP of Lucifer Yellow filled neurons was -33.6 mV; mean RMP of HRP filled neurons was 42.3 mV). In contrast, the mean RMPs of Lucifer Yellow and HRP marked glia was -68 Mv and -75 mV respectively, and the quality of "staining" appeared to be more closely related to the RMP.
Oxygen radicals mediate cell inactivation by acridine dyes, fluorescein, and Lucifer Yellow CH
Photochem Photobiol 1987 Jul;46(1):45-53.PMID:3039547DOI:10.1111/j.1751-1097.1987.tb04734.x.
Acridine dyes, fluorescein and Lucifer Yellow CH are fluorescent photosensitizers used experimentally to selectively stain and photodynamically destroy eukaryotic cells and subcellular structures. We have determined that the mechanism of light- and oxygen-dependent inactivation of E. coli by these dyes involves oxygen radicals and hydrogen peroxide. All of the dyes oxidized NAD(P)H+ under illumination. Superoxide (O2), detected as the superoxide dismutase (SOD)-inhibitable reduction of ferricytochrome c, was a major product of the dye sensitized photooxidation. Cationic acridine dyes penetrated the membranes of E. coli and were photoreduced intracellularly. Reduced dyes diffused back into the medium and mediated the reduction of extracellular ferricytochrome c. The anionic dyes fluorescein and Lucifer Yellow CH were unable to mediate extracellular cytochrome c reduction, indicating that these dyes were impermeable to the E. coli membrane. Acridine dyes, when illuminated, inhibited the growth of E. coli in a rich medium, and induced the synthesis of SOD. Fluorescein and Lucifer Yellow CH did not inhibit growth or induce SOD synthesis because they were unable to enter the cells. Superoxide (O2) and hydrogen peroxide (H2O2), generated by the enzyme xanthine oxidase were toxic to E. coli B. Inactivation by xanthine oxidase was partially inhibited by exogenous SOD and completely inhibited by exogenous catalase or SOD plus catalase. Similarly, exogenous SOD plus catalase protected against inactivation by acridines and fluorescein-NADH or lucifer yellow CH-NADH mixtures. Prior induction of superoxide dismutase and catalase in E. coli B significantly protected cells against a subsequent challenge by illuminated acridine dyes. SOD and catalases preinduction combined with additions of exogenous SOD and catalase completely protected E. coli B against photodynamic inactivation by acridine yellow. The hydroxyl radical scavengers, dimethyl sulfoxide, sodium benzoate and thiourea, protected E. coli B against photodynamic inactivation by acridine orange. The results implicate O2, H2O2, and the hydroxyl radical (OH) as underlying molecular agents of the phototoxicity mediated by acridine orange, acridine yellow, fluorescein and Lucifer Yellow CH.
Microinjection of Lucifer Yellow CH into sea urchin eggs and embryos
Cell Tissue Res 1983;234(2):309-18.PMID:6640624DOI:10.1007/BF00213770.
Eggs and embryos of Arbacia punctulata were microinjected with the fluorescent dye, Lucifer Yellow CH, using a simple pressure injection system. When injected into eggs that were subsequently fertilized, the dye was distributed throughout all cells of the developing embryo. If one cell of a two-cell embryo was injected, dye did not diffuse into the uninjected blastomere. During subsequent development, all progeny of the injected cell contained dye resulting in an embryo that was half-fluorescent. Blue light irradiation of a two-cell embryo, one cell of which had been injected with Lucifer yellow, caused the injected blastomere to stop further divisions while the uninjected blastomere developed normally and was free of dye. These results indicate that the first two blastomeres of Arbacia embryos are not electrically coupled, nor up to the time of hatching, is there any coupling between cells in one half of the first cleavage plane and cells in the other half.
Effect of superoxide derived from Lucifer Yellow CH on voltage-gated currents of mouse taste bud cells
Chem Senses 2008 Jun;33(5):425-32.PMID:18319263DOI:10.1093/chemse/bjn009.
Lucifer Yellow CH (LY), a fluorescent membrane-impermeable cell marker dye, has been routinely loaded into cells through recording electrodes to visualize these cells after electrophysiological investigation, without considering its pharmacological effect. Recently, we showed that the exposure of cells loaded with LY to light for microscopy produced unidentified radical species that retarded the inactivation of voltage-gated Na+ currents irreversibly (Higure Y et al. 2003). Here, we show that superoxide dismutase, an enzyme that decomposes superoxide, reverses the retardation effect, which assures that superoxide is the unidentified radical species. The estimated mean lifetime of superoxide in recording electrodes (in the absence of cytoplasm) is approximately 6 min, and hence, the Na+ currents are retarded even in the dark, when LY is exposed to light before being loaded into the cell. Superoxide has no effect on voltage-gated Cl- currents. These results show that superoxide action on ion channels is rather selective. The breakdown of superoxide inside cells and the effect of endogenous superoxide on the superoxide-susceptible channels are discussed.
Lucifer yellow, neuronal marking and paraffin sectioning
Stain Technol 1983 Jul;58(4):189-92.PMID:6197772DOI:10.3109/10520298309066784.
An unusual procedure with the dye lucifer yellow has provided stable neuronal marking that survives paraffin embedding and sectioning. Lucifer Yellow CH was dissolved in an electrolyte containing formaldehyde and injected into the large interneurons of a cricket. Intense fluorescence in the axoplasm was retained even after conventional histological procedures.