Fluo-3 (sodium salt)
目录号 : GC43683A fluorescent calcium indicator
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >90.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Fluo-3 is a fluorescent calcium indicator commonly used in flow cytometry and cell-based experiments to detect changes in intracellular calcium levels. Its absorption maximum at 506 nm makes it compatible with excitation at 488 nm by argon-ion laser sources. Fluo-3 provides intense fluorescence upon binding calcium, detected at a maximum emission at 526 nm which can be monitored by FL1 (green, 525 nm band pass) sensors in flow cytometry.
| Cas No. | SDF | ||
| Canonical SMILES | O=C1C(Cl)=CC(C(O2)=C1)=C(C3=CC=C(N(CC([O-])=O)CC([O-])=O)C(OCCOC4=C(N(CC([O-])=O)CC([O-])=O)C=CC(C)=C4)=C3)C5=C2C=C([O-])C(Cl)=C5.[Na+].[Na+].[Na+].[Na+].[Na+] | ||
| 分子式 | C36H25Cl2N2O13•5Na | 分子量 | 879.4 |
| 溶解度 | Water: soluble | 储存条件 | Store at -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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.1371 mL | 5.6857 mL | 11.3714 mL |
| 5 mM | 0.2274 mL | 1.1371 mL | 2.2743 mL |
| 10 mM | 0.1137 mL | 0.5686 mL | 1.1371 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。
NCX3 alleviates ethanol-induced apoptosis of SK-N-SH cells via the elimination of intracellular calcium ions
Toxicol In Vitro 2021 Apr;72:105104.PMID:33516933DOI:10.1016/j.tiv.2021.105104.
Long-term alcohol intake may cause nerve cell apoptosis and induce various encephalopathies. Previously, we have shown that the expression of Na+/Ca2+ exchanger 3 (NCX3) was associated with the intracellular calcium concentration ([Ca2+]i) and apoptosis, involved in the spatial memory impairment in male C57BL/6 mice with chronic ethanol (EtOH) exposure. However, the mechanism involved is unclear. Here, we investigated the expression of NCX3 and its protective effect on SK-N-SH cells (a nerve cell line) after EtOH exposure. [Ca2+]i was measured using Fluo-3 AM reagent. Cell viability and the apoptotic rate were assayed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) and flow cytometry, respectively. The expression of p-cAMP-responsive element binding protein1(p-CREB 1), NCX3 protein, and mRNA were observed using Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. Cleaved-caspase-3, caspase-3, rabbit anti- poly (ADP-ribose) polymerase-1 (PARP-1) and calpain-1 proteins were used to assess the degree of apoptosis. Our results showed that EtOH increased [Ca2+]i and apoptosis of SK-N-SH cells in a concentration- and time-dependent manner. The expression of NCX3 protein and mRNA was up regulated obviously after SK-N-SH cells were treated with EtOH. The phosphorylation levels of Akt and CREB 1 were up regulated in cells treated with EtOH. The expression of NCX3 protein was reduced in the SK-N-SH cells treated with Akt phosphorylation inhibitor (LY294002). The [Ca2+]i and apoptosis rate of SK-N-SH cells increased 1.31-fold and 1.52-fold after silencing NCX3 compared with those treated with 200 mM EtOH alone for 2 d. In contrast, the [Ca2+]i and apoptosis rate of SK-N-SH cells decreased 0.26-fold and 0.35-fold after overexpression of NCX3 in the 2 d-200 mM EtOH treatment group. These results suggest that NCX3 plays a critical role in neuronal protection via the elimination of intracellular Ca2+, which may be a promising target for the prevention and treatment of encephalopathy after ethanol exposure.
Ca2+ sparks induced by Na/Ca exchange
Cell Calcium 2003 Jul;34(1):11-7.PMID:12767888DOI:10.1016/s0143-4160(03)00017-4.
Whether Ca(2+) influx on the Na/Ca exchanger (NCX) can trigger elementary sarcoplasmic reticulum (SR) Ca(2+) release events (Ca(2+) sparks) is controversial. We imaged [Ca(2+)](i) (Nipkow confocal microscope and Fluo-3) in left ventricular myocytes isolated from wild type (WT) and transgenic (TG) mice overexpressing NCX 2.5-fold. Sudden activation of Ca(2+) influx via NCX induced by abrupt exposure to "0" [Na(+)](o)/normal [Ca(2+)](o) solution by means of a rapid solution switcher-induced Ca(2+) sparks in NCX TG myocytes in 425+/-17 ms, n=21. The diameter and amplitude (F/F(0)) of these sparks (2.74+/-0.14 microm, F/F(0)=2.16+/-0.06, n=18) were similar to those induced by field stimulation of myocytes in the presence of 20 microM nifedipine (2.70+/-0.10 microm, F/F(0)=1.98+/-0.08, n=17). In WT myocytes no Ca(2+) sparks were observed within the first 600 ms after abrupt removal of extracellular Na. In parallel experiments, voltage clamp current measurements (-80 mV) showed that the Na/Ca exchange current (I(NCX)) began within 60 ms of activation of the switcher, and peaked at 312+/-57 pA in TG myocytes within 300-500 ms. I(Ca,L) in 20 microM nifedipine was 10.3+/-4.3 pA, n=7. These results indicate that Ca(2+) entering the myocyte via NCX can cause Ca(2+) sparks which are similar to those elicited by electrical stimulation. However, Ca(2+) influx on NCX is much less efficient in inducing Ca(2+) sparks than Ca(2+) influx via I(Ca,L).
Cryptococcus neoformans Ca(2+) homeostasis requires a chloride channel/antiporter Clc1 in JEC21, but not in H99
FEMS Yeast Res 2012 Feb;12(1):69-77.PMID:22093100DOI:10.1111/j.1567-1364.2011.00763.x.
CLC-type chloride/proton antiporters are required for copper/iron homeostasis in fungi. A relationship between CLCs and Ca(2+) homeostasis has not been found before. Here we demonstrate the requirement of the antiporter CLC1 for Ca(2+) homeostasis/signaling in Cryptococcus neoformans. The deletion of CLC1 in JEC21 resulted in a mutant hypersensitive to cyclosporine A, an inhibitor of calcineurin. Intracellular Ca(2+) deficiency in the mutant Tx1 was confirmed with Fluo-3 staining epi-fluorescence microscopy. Tx1 failed to grow at elevated temperature and in SDS and displayed defects in cell wall integrity and cell separation. This defective phenotype is because of Ca(2+) deficiency that was restorable by exogenous Ca(2+) . In contrast, H99 CLC1 was dispensable for Ca(2+) homeostasis and had no comparable defective consequences if deleted, suggesting divergent roles of CLCs in Ca(2+) homeostasis. Distinct Ca(2+) homeostasis mechanisms may contribute the virulence difference between the two strains. This work reveals a novel action of CLC antiporters in fungi and may provide information as to the evolution of pathogenicity among cryptococcal strains.
Cytosolic calcium concentration is reduced by photolysis of a nitrosyl ruthenium complex in vascular smooth muscle cells
Nitric Oxide 2006 Nov;15(3):252-8.PMID:16564714DOI:10.1016/j.niox.2006.02.001.
The effect of the NO donors cis-[RuCl(bpy)(2)(NO)](PF(6)) (RUNOCL) and sodium nitroprusside (SNP) on the cytosolic Ca(2+) concentration ([Ca(2+)](c)) was studied in cells isolated from the rat aorta smooth muscle of cells isolated from the rat aorta smooth muscle. SNP is a metal nitrosyl complex made up of iron, cyanide groups, and a nitro moiety; the RUNOCL complex is made up of ruthenium and bipyridine ligands, with chloride and nitrosyl groups in the ruthenium axial positions. Rat aorta smooth muscle cells were loaded with Fluo-3 acetoxymethyl ester (Fluo-3 AM) and imaged by a confocal scanning laser microscope excited with the 488 nm line of the argon ion laser. Fluorescence emission was measured at 510 nm. One of the NO donors, RUNOCL (100 micromol/L) or SNP (100 micromol/L), was then added to the cell chamber and the fluorescent intensity percentage (%IF) was measured after 240 s. RUNOCL reduced the %IF to 60.0+/-10.0% of the initial value. After treatment with the soluble guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ) (10 micromol/L), the measurement of %IF was 81.0+/-5.0% (n=4). In the presence of tetraethylammonium (TEA) (1 mmol/L) the %IF was 79.0+/-6.4% (n=4). A combination of ODQ and TEA increased the %IF to 97.0+/-3.5% (n=4). As for SNP, it reduced the %IF to 81.4+/-4.7% (n=4), but this effect was inhibited by ODQ (%IF 94.0+/-3.6%; n=4) and TEA (%IF 88.0+/-2.1%; n=4). The combination of ODQ and TEA increased (%IF 92.0+/-2.8%; n=4). Taken together, these results indicate that both the new NO donor RUNOCL and SNP reduce [Ca(2+)](c). Our data also give evidence that soluble guanylyl cyclase and K(+) channels sensitive to TEA are involved in the mechanisms responsible for the reduction in [Ca(2+)](c) of the rat aorta smooth muscle cells.
Endothelium-independent vasorelaxant effect of sodium ferulate on rat thoracic aorta
Life Sci 2009 Jan 16;84(3-4):81-8.PMID:19038273DOI:10.1016/j.lfs.2008.11.003.
Aims: This study was designed to investigate the effects of sodium ferulate (SF) on rat isolated thoracic aortas and the possible mechanisms. Main methods: Isometric tension was recorded in response to drugs in organ bath. Cytosolic free Ca(2+) concentration ([Ca(2+)](i)) was measured using Fluo-3 in cultured rat aortic smooth muscle cells (RASMC). Key findings: SF (0.1-30 mM) relaxed the isolated aortic rings precontracted with phenylephrine (PE) and high-K(+) in a concentration-dependent manner with respective pD(2) of 2.7+/-0.02 and 2.6+/-0.06. Mechanical removal of endothelium did not significantly modify the SF-induced relaxation. In Ca(2+)-free solution, SF noticeably inhibited extracellular Ca(2+)-induced contraction in high-K(+) and PE pre-challenged rings, and suppressed the transient contraction induced by PE and caffeine. The vasorelaxant effect of SF was unaffected by various K(+) channel blockers such as tetraethylammonium, glibenclamide, 4-aminopyridine, and barium chloride. In addition, SF concentration-dependently reduced the contraction induced by phorbol-12-myristate-13-acetate, an activator of protein kinase C (PKC), in the absence of extracellular Ca(2+), with the pD(2) of 2.9+/-0.03. In RASMC, SF had no effect on PE- or KCl-induced [Ca(2+)](i) increase either in the presence or in the absence of external Ca(2+). Significance: These results indicate that SF acts directly as a non-selective relaxant to vascular smooth muscle. The direct inhibition of the common pathway after [Ca(2+)](i) increase may account for the SF-induced relaxation in Ca(2+)-dependent contraction, while the blockage of the PKC-mediated contractile mechanism is likely responsible for the SF-induced relaxation in Ca(2+)-independent contraction.