| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Cell membrane lipid bilayer and transmembrane voltage. Di-8-ANEPPS is a voltage-sensitive fluorescent dye that incorporates into cell membranes and exhibits changes in fluorescence in response to membrane potential variations. The dye is not fluorescent in water, but when bound to the lipid bilayer, it becomes strongly fluorescent. Its fluorescence excitation and emission spectra change in response to changes in voltage, with an optical response fast enough to detect transient membrane potential changes in excitable cells.
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| ln Vitro |
Mouse cells stained with Di-8-ANEPPS were exposed to voltage clamp pulses and submerged in a solution containing Na+. The Di-8-ANEPPS signal generally reflects the system voltage, with an asymmetric positive component when depolarizing pulses are delivered. This is thought to reflect propagating action potentials escaping from the voltage-clamped plasma membrane. Its temporal features are no different from those of the "center" signal. It is worth noticing that the signal peak comes 0.3 milliseconds following the leading edge of the depolarizing pulse [1].
In vitro, Di-8-ANEPPS is used to noninvasively measure transmembrane voltage in live cells. The dye exhibits a membrane potential-dependent shift in excitation spectra, with a fluorescence change of 2-10% per 100 mV. It is widely used for monitoring fast membrane potential changes in excitable cells such as neurons and cardiomyocytes. The Di-8-ANEPPS signal generally reflects the system voltage, with an asymmetric positive component when depolarizing pulses are delivered. Its longer carbon chain tails make it more stable than Di-4-ANEPPS for long-duration experiments. |
| ln Vivo |
In vivo, Di-8-ANEPPS is used to monitor membrane potential in tissues and whole organisms. The dye can be applied to exposed tissues or loaded into cells for in vivo imaging of electrical activity. It is used in studies of cardiac electrophysiology, neuronal activity, and other physiological processes that involve changes in membrane potential. Its fast response kinetics make it suitable for detecting action potentials and other transient electrical events.
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| Enzyme Assay |
In vitro fluorescence assays for Di-8-ANEPPS measure voltage-dependent fluorescence changes. The dye is dissolved in DMSO or ethanol and added to cell cultures or membrane preparations at concentrations of 1-10 microM. Cells are excited at 467 nm, and emission is measured at 631 nm. Changes in membrane potential are induced by adding ionophores (e.g., valinomycin) or by electrical stimulation. The fluorescence signal is recorded using a fluorescence microscope, plate reader, or electrophysiology setup.
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| Cell Assay |
In vitro cell-based assays using Di-8-ANEPPS are performed with cultured neurons, cardiomyocytes, or other excitable cells. Cells are loaded with the dye by incubation in buffer containing 1-10 microM Di-8-ANEPPS for 15-30 minutes at 37degC. After washing, cells are placed on a microscope stage and stimulated electrically or chemically. Fluorescence changes are recorded using high-speed cameras or photomultiplier tubes. The dye's fast response kinetics allow detection of action potentials and other transient events.
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| Animal Protocol |
In vivo animal studies with Di-8-ANEPPS are conducted in various model organisms. The dye is applied topically to exposed tissues (e.g., heart, brain) or injected intravenously. Optical mapping of membrane potential is performed using high-speed cameras and appropriate excitation/emission filters. The dye is used to study cardiac arrhythmias, neuronal network activity, and other electrophysiological phenomena.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of Di-8-ANEPPS are characteristic of lipophilic fluorescent dyes. With a molecular weight of 592.9 and a molecular formula of C36H52N2O3S, the compound is soluble in DMF (2 mg/mL), DMSO (2 mg/mL), and ethanol (100 microg/mL). It is stored as a crystalline solid at -20degC. The dye has excitation maxima at 208, 253, 320, and 501 nm and emission at 631 nm. As a research dye, pharmacokinetic studies are not typically performed.
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| Toxicity/Toxicokinetics |
Di-8-ANEPPS is intended for research use only and is not for human therapeutic use. Standard safety precautions for handling chemical compounds apply. The compound is not approved for clinical use.
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| References | |
| Additional Infomation |
Di-8-ANEPPS is a research-grade voltage-sensitive fluorescent dye used to monitor membrane potential in live cells and tissues. It is a naphthylstyrene-based dye with a molecular formula of C36H52N2O3S and a molecular weight of 592.9. The dye exhibits excitation/emission maxima of approximately 467/631 nm when bound to lipid bilayers. It is widely used for monitoring fast membrane potential changes in excitable cells. Not approved for clinical use.
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| Molecular Formula |
C36H52N2O3S
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|---|---|
| Molecular Weight |
592.87468
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| Exact Mass |
592.37
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| CAS # |
157134-53-7
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| PubChem CID |
6508718
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| Appearance |
Brown to red solid powder
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| LogP |
9.841
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
20
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| Heavy Atom Count |
42
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| Complexity |
789
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCCCCCCCN(CCCCCCCC)C1=CC2=C(C=C1)C=C(C=C2)/C=C/C3=CC=[N+](C=C3)CCCS(=O)(=O)[O-]
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| InChi Key |
IXFSUSNUALIXLU-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C36H52N2O3S/c1-3-5-7-9-11-13-25-38(26-14-12-10-8-6-4-2)36-21-20-34-30-33(18-19-35(34)31-36)17-16-32-22-27-37(28-23-32)24-15-29-42(39,40)41/h16-23,27-28,30-31H,3-15,24-26,29H2,1-2H3
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| Chemical Name |
3-[4-[(E)-2-[6-(dioctylamino)naphthalen-2-yl]ethenyl]pyridin-1-ium-1-yl]propane-1-sulfonate
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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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| 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 : ~1 mg/mL (~1.69 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 | 1.6867 mL | 8.4336 mL | 16.8671 mL | |
| 5 mM | 0.3373 mL | 1.6867 mL | 3.3734 mL | |
| 10 mM | 0.1687 mL | 0.8434 mL | 1.6867 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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