Fluo-2 AM
(Synonyms: 钙离子荧光探针FLUO-2AM) 目录号 : GC49458A cell-permeable fluorescent calcium indicator
Cas No.:1070771-36-6
Sample solution is provided at 25 µL, 10mM.
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- Purity: >95.00%
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Fluo-2 AM is a cell-permeable fluorescent calcium indicator.1,2 It has been used to measure the activity of the neuropeptide Y4 receptor, as well as to detect intracellular calcium in tetrandine-stimulated primary rabbit corpus cavernosum smooth muscle cells. It binds to calcium (Kd = 290 nM) and displays excitation/emission maxima of 490/515 nm, respectively. Fluo-2 AM is also available in a cell-impermeable form .
1.Sliwoski, G., Schubert, M., Stichel, J., et al.Discovery of small-molecule modulators of the human Y4 receptorPLoS One11(6)e0157146(2016) 2.Liu, J.-H., Chen, J., Wang, T., et al.Effects of tetrandrine on cytosolic free calcium concentration in corpus cavernosum smooth muscle cells of rabbitsAsian J. Androl.8(4)405-409(2006)
Cas No. | 1070771-36-6 | SDF | Download SDF |
别名 | 钙离子荧光探针FLUO-2AM | ||
Canonical SMILES | O=C1C=CC2=C(C3=CC(OCCOC4=CC(C)=CC=C4N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)=C(N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)C=C3)C5=C(C=C(C=C5)OCOC(C)=O)OC2=C1 | ||
分子式 | C51H52N2O23 | 分子量 | 1061 |
溶解度 | N/A | 储存条件 | -20°C |
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 | 0.9425 mL | 4.7125 mL | 9.4251 mL |
5 mM | 0.1885 mL | 0.9425 mL | 1.885 mL |
10 mM | 0.0943 mL | 0.4713 mL | 0.9425 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。
Rhythmic Calcium Events in the Lamina Propria Network of the Urinary Bladder of Rat Pups
Front Syst Neurosci 2017 Dec 11;11:87.PMID:29321730DOI:10.3389/fnsys.2017.00087.
The lamina propria contains a dense network of cells, including interstitial cells (ICs), that may play a role in bladder function by modulating communication between urothelium, nerve fibers and smooth muscle or acting as pacemakers. Transient receptor potential vanilloid 4 (TRPV4) channels allow cation influx and may be involved in sensing stretch or chemical irritation in urinary bladder. Urothelium was removed from rats (P0-Adult), cut into strips, and loaded with a Ca2+ fluorescent dye (Fluo-2 AM leak resistant or Cal 520) for 90 min (35-37°C) to measure Ca2+ events. Ca2+ events were recorded for a period of 60 seconds (s) in control and after drug treatment. A heterogeneous network of cells was identified at the interface of the urothelium and lamina propria of postnatal rat pups, aged ≤ postnatal (P) day 21, with diverse morphology (round, fusiform, stellate with numerous projections) and expressing platelet-derived growth factor receptor alpha (PDGFRα)- and TRPV4-immunoreactivity (IR). Ca2+ transients occurred at a slow frequency with an average interval of 30 ± 8.6 s. Waveform analyses of Ca2+ transients in cells in the lamina propria network revealed long duration Ca2+ events with slow upstrokes. We observed slow propagating waves of activity in the lamina propria network that displayed varying degrees of coupling. Application of the TRPV4 agonist, GSK1016790 (100 nM), increased the duration of Ca2+ events, the number of cells with Ca2+ events and the integrated Ca2+ activity corresponding to propagation of activity among cells in the lamina propria network. However, GSK2193874 (1 μM), a potent antagonist of TRPV4 channels, was without effect. ATP (1 μM) perfusion increased the number of cells in the lamina propria exhibiting Ca2+ events and produced tightly coupled network activity. These findings indicate that ATP and TRPV4 can activate cells in the laminar propria network, leading to the appearance of organized propagating wavefronts.
A Portulaca oleracea L. extract promotes insulin secretion via a K+ATP channel dependent pathway in INS-1 pancreatic β-cells
Nutr Res Pract 2018 Jun;12(3):183-190.PMID:29854323DOI:10.4162/nrp.2018.12.3.183.
Background/objective: This study was designed to investigate how a Portulaca oleracea L. extract (POE) stimulates insulin secretion in INS-1 pancreatic β-cells. Materials/method: INS-1 pancreatic β-cells were incubated in the presence of various glucose concentrations: 1.1 or 5.6, 16.7 mM glucose. The cells were treated with insulin secretagogues or insulin secretion inhibitor for insulin secretion assay using an insulin ELISA kit. In order to quantify intracellular influx of Ca2+ caused by POE treatment, the effect of POE on intracellular Ca2+ in INS-1 pancreatic β-cells was examined using Fluo-2 AM dye. Results: POE at 10 to 200 µg/mL significantly increased insulin secretion dose-dependently as compared to the control. Experiments at three glucose concentrations (1.1, 5.6, and 16.7 mM) confirmed that POE significantly stimulated insulin secretion on its own as well as in a glucose-dependent manner. POE also exerted synergistic effects on insulin secretion with secretagogues, such as L-alanine, 3-isobutyl-1-methylxanthine, and especially tolbutamide, and at a depolarizing concentration of KCl. The insulin secretion caused by POE was significantly attenuated by treatment with diazoxide, an opener of the K+ATP channel (blocking insulin secretion) and by verapamil (a Ca2+ channel blocker). The insulinotropic effect of POE was not observed under Ca2+-free conditions in INS-1 pancreatic β-cells. When the cells were preincubated with a Ca2+ fluorescent dye, Fluo-2 (acetoxymethyl ester), the cells treated with POE showed changes in fluorescence in red, green, and blue tones, indicating a significant increase in intracellular Ca2+, which closely correlated with increases in the levels of insulin secretion. Conclusions: These findings indicate that POE stimulates insulin secretion via a K+ATP channel-dependent pathway in INS-1 pancreatic β-cells.
Scopoletin stimulates the secretion of insulin via a KATP channel-dependent pathway in INS-1 pancreatic β cells
J Pharm Pharmacol 2022 Sep 1;74(9):1274-1281.PMID:35099527DOI:10.1093/jpp/rgab143.
Objectives: In this study, we investigated whether scopoletin stimulated the secretion of insulin in pancreatic β cells as well as the underlying mechanism involved in this process. Methods: We incubated the INS-1 pancreatic β cells with various concentrations of glucose (1.1, 5.6 or 16.7 mM) in the presence or absence of scopoletin. We then analysed the secretion of insulin in the cells treated with insulin secretion inhibitors or secretagogues. The intracellular influx of calcium induced by scopoletin was also analysed using the Fluo-2 AM dye. Key findings: We found that scopoletin (1-20 µM) markedly induced the secretion of insulin in a glucose concentration-dependent manner compared with the control. At depolarizing concentrations of potassium chloride (KCl), scopoletin markedly enhanced the insulin secretion compared with the cells which were treated only with KCl. Moreover, the treatment with diazoxide-opening K+ATP channel and verapamil blocking Ca2+ channel significantly decreased the scopoletin-induced increase in insulin secretion. After the pre-treatment of cells with a Ca2+ fluorescent dye, treatment with 20 µM scopoletin resulted in a significant increase in the influx of intracellular Ca2+, exhibiting fluorescence changes in various spectra. Conclusions: Scopoletin stimulates the secretion of insulin via a K+ATP channel-dependent pathway in the INS-1 pancreatic β cells.