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| Targets |
FM1-84 targets cell membranes, specifically the lipid bilayer. As a styryl pyridinium dye, the compound partitions into membranes and exhibits fluorescence enhancement upon membrane binding. The dye's fluorescence properties are sensitive to membrane potential, lipid packing, and membrane curvature, making it a useful tool for studying membrane dynamics. In neurobiology, FM dyes are used to label synaptic vesicles and monitor synaptic activity through the uptake and release of the dye during endocytosis and exocytosis. The compound does not target a specific protein but rather the lipid environment of membranes.
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| ln Vitro |
In vitro, FM1-84 is used as a fluorescent membrane probe to label cell membranes and study membrane dynamics. The dye exhibits fluorescence enhancement upon binding to membranes, allowing visualization of membrane structures by fluorescence microscopy. In neuronal cultures, FM1-84 is used to label synaptic vesicles and monitor synaptic activity. The dye is taken up into synaptic vesicles during endocytosis and released during exocytosis, providing a readout of synaptic transmission. The compound's spectral properties (excitation and emission wavelengths) determine its suitability for specific imaging applications.
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| ln Vivo |
In vivo, FM1-84 can be used for imaging studies in animal models, although its primary application is in vitro and ex vivo. The dye can be injected into tissues or animals to label membranes and study membrane dynamics in living systems. However, its use in vivo may be limited by tissue penetration, photobleaching, and clearance. The compound is primarily used as a research tool for studying membrane biology and synaptic function.
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| Enzyme Assay |
In vitro assays with FM1-84 involve incubating cells or membranes with the dye and measuring fluorescence by spectroscopy or microscopy. The dye's fluorescence enhancement upon membrane binding can be quantified to assess membrane binding or to monitor membrane dynamics. In synaptic vesicle labeling studies, neurons are incubated with FM1-84, and dye uptake and release are monitored by fluorescence microscopy or flow cytometry. These assays are used to study endocytosis, exocytosis, and synaptic transmission.
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| Cell Assay |
Cell-based assays with FM1-84 are conducted in cultured cells, particularly neurons. Cells are incubated with the dye at various concentrations and time points. Fluorescence is measured by microscopy or flow cytometry. In synaptic vesicle labeling studies, neurons are stimulated to induce endocytosis and uptake of the dye, followed by stimulation to induce exocytosis and release of the dye. The change in fluorescence provides a measure of synaptic activity. These assays are used to study synaptic function and plasticity.
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| Animal Protocol |
In vivo animal protocols for FM1-84 are limited, as the compound is primarily used in vitro and ex vivo. For in vivo imaging, the dye may be injected into tissues or animals, and fluorescence is monitored by imaging techniques. However, specific protocols are not extensively documented. Researchers should consult primary literature for specific applications.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of FM1-84 have not been extensively characterized. The compound is a fluorescent dye used as a research tool, not as a therapeutic agent. Its physicochemical properties include molecular weight and spectral properties (excitation and emission wavelengths). The compound is typically dissolved in DMSO or aqueous buffers for experiments. Storage: protect from light.
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| Toxicity/Toxicokinetics |
Safety and toxicology data for FM1-84 are limited. As a fluorescent dye, the compound may have potential toxicity at high concentrations or prolonged exposure. Standard laboratory safety precautions should be followed when handling the compound. The compound should be protected from light and stored properly. The compound is for research use only and is not for human use.
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| References | |
| Additional Infomation |
FM1-84 has CAS number 303727-66-4 and is a fluorescent membrane probe of the styryl pyridinium class. It is used for studying membrane dynamics, synaptic vesicle trafficking, and endocytosis/exocytosis. The dye exhibits fluorescence enhancement upon membrane binding. Purity is typically ≥95%. Protect from light. Not for human use; for research purposes only.
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| Molecular Formula |
C32H53N3.2BR
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|---|---|
| Molecular Weight |
479.78332
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| Exact Mass |
479.422
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| CAS # |
303727-66-4
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| PubChem CID |
57463566
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| Appearance |
Brown to reddish brown ointment
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| LogP |
2.49
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
18
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| Heavy Atom Count |
37
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| Complexity |
507
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N(C1C=CC(C=CC2=CC=[N+](CCC[N+](CC)(CC)CC)C=C2)=CC=1)(CCCCC)CCCCC.[Br-]
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| InChi Key |
QCVUFCBGGWOMSS-UHFFFAOYSA-L
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| InChi Code |
InChI=1S/C32H53N3.2BrH/c1-6-11-13-25-34(26-14-12-7-2)32-20-18-30(19-21-32)16-17-31-22-27-33(28-23-31)24-15-29-35(8-3,9-4)10-5;;/h16-23,27-28H,6-15,24-26,29H2,1-5H3;2*1H/q+2;;/p-2
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| Chemical Name |
3-[4-[(E)-2-[4-(dipentylamino)phenyl]ethenyl]pyridin-1-ium-1-yl]propyl-triethylazanium;dibromide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ≥ 125 mg/mL (~195.44 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0843 mL | 10.4214 mL | 20.8429 mL | |
| 5 mM | 0.4169 mL | 2.0843 mL | 4.1686 mL | |
| 10 mM | 0.2084 mL | 1.0421 mL | 2.0843 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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