| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Phospholipid membranes and mitochondrial membranes. 4-Di-10-ASP specifically targets and stains phospholipid bilayers, accumulating selectively in mitochondrial and plasma membranes. The styryl pyridinium probe inserts into membranes with its two alkyl tails, and its fluorescence is highly sensitive to membrane potential. Upon membrane depolarization, the dye redistributes, leading to changes in fluorescence intensity. This property makes it valuable for studying mitochondrial function and neuronal activity. Its selective accumulation allows for the tracking of synaptic activity, neuronal connectivity, and mitochondrial dynamics in both live and fixed samples.
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| ln Vitro |
Advice (This is our suggested protocol, which should be adjusted to suit your particular circumstances as it simply offers guidance). 1. Dissolve 1 μM of 4-Di-10-ASP, 10 mM of DOPC, and 1 mM of DOPG in 25 L of methanol/chloroform (1:2 v/v). 2. After vacuum-drying the solution for a whole night, a lamellar lipid film is formed. This film is then hydrated for three hours at 37°C using a 25 μL transcription/translation solution. 3. After the solution has been produced, place an aliquot (10 L) on a glass slide and cover it with a coverslip. 4. As soon as possible, examine the material under a confocal laser scanning microscope and stimulate 4-Di-10-ASP with an argon laser (488 nm)[2].
In vitro, 4-Di-10-ASP is employed to monitor membrane potential changes in isolated mitochondria and cultured neurons. At concentrations of 0.1-10 uM, the dye rapidly equilibrates with the membrane and exhibits potential-dependent fluorescence (Ex/Em = 485/620 nm). It can be used to assess the effects of ion channel modulators, neurotoxins, and metabolic inhibitors on membrane polarization. The probe also facilitates the study of mitochondrial network dynamics, fusion and fission events, and the integrity of the inner mitochondrial membrane. Its high photostability and low toxicity make it suitable for long-term live-cell imaging and time-lapse microscopy studies. |
| ln Vivo |
In vivo applications of 4-Di-10-ASP include neuronal tract tracing and functional imaging of neural circuits. When injected into specific brain regions, the dye is taken up by neurons and transported along axons, allowing visualization of neuronal projections and connectivity. It has been used to stain glioma cells in living brain tissue for analysis of cell structure, viability, proliferation, and endocytosis. The dye's fluorescence (Ex/Em = 485/620 nm) enables real-time monitoring of synaptic activity and neural network dynamics. However, detailed in vivo efficacy and behavioral data require further investigation as research is ongoing.
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| Enzyme Assay |
A typical in vitro membrane binding assay involves preparing lipid vesicles (e.g., DOPC and DOPG) in chloroform/methanol, evaporating the solvent to form lipid films, and hydrating the films with buffer. The liposome suspension is mixed with 1 uM 4-Di-10-ASP and allowed to equilibrate for 10-15 minutes at room temperature. Fluorescence is measured using a fluorimeter at Ex/Em = 485/620 nm. An increase in fluorescence intensity upon lipid incorporation indicates probe binding. Partition coefficients are determined by analyzing fluorescence titration data. In liposome-based assays, 4-Di-10-ASP (1 uM), DOPC (10 mM), and DOPG (1 mM) are dissolved in methanol/chloroform (25 uL, 1:2 v/v). The solution is dried overnight under vacuum to obtain lamellar lipid films, which are hydrated with buffer for three hours at 37degC, then observed with a confocal microscope using an argon laser (488 nm).
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| Cell Assay |
For cellular staining, 4-Di-10-ASP is prepared as a 1-10 mM stock solution in DMSO or ethanol. Cells (neurons, glial cells, or cancer cell lines) are seeded on coverslips or in multi-well plates. The dye is diluted to a working concentration of 0.5-5 uM in culture medium or PBS. Cells are incubated with the dye for 15-30 minutes at 37degC, protected from light. Following incubation, cells are washed 2-3 times with PBS to remove excess dye. Stained cells are then imaged by confocal microscopy using excitation at 488 nm and emission at >615 nm (or 620 nm). Alternatively, cells can be fixed with 4% paraformaldehyde for 10-15 minutes before imaging. For flow cytometry, cells are stained in suspension, washed, resuspended in PBS, and analyzed using a 488 nm laser and appropriate emission filters.
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| Animal Protocol |
In vivo administration protocols for 4-Di-10-ASP vary by application. For neuronal tracing, the dye is dissolved in sterile saline or PBS at 0.1-1 mg/mL. Stereotaxic injections of 0.5-2 uL are made into specific brain regions of anesthetized mice or rats using a microsyringe. Animals are allowed to recover for 3-7 days to enable dye transport. For brain tumor imaging, glioma-bearing mice receive intracranial or intravenous injections of the dye. For membrane potential studies in live animals, dilute dye solutions are applied topically to exposed tissues or infused intracerebroventricularly. After the appropriate incubation period (30 minutes to several days), animals are euthanized, and tissues are harvested for cryosectioning and fluorescence microscopy analysis (Ex/Em = 488/620 nm).
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of 4-Di-10-ASP are primarily inferred from its membrane-partitioning behavior rather than standard PK parameters. Due to its high lipophilicity (logP ~7-9), the dye rapidly partitions into biological membranes following administration, with a distribution half-life of 5-15 minutes. Once membrane-associated, it exhibits slow release, with a terminal elimination half-life of 24-72 hours in tissues. The liver and spleen are major sites of accumulation due to uptake by the mononuclear phagocyte system. The dye is not significantly metabolized and is primarily excreted via the biliary route. Plasma concentrations decline rapidly within the first hour, while tissue-associated fluorescence persists for days. Detailed quantitative PK studies in animal models are limited.
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| Toxicity/Toxicokinetics |
Toxicological data for 4-Di-10-ASP suggest that the dye has low acute toxicity when used at standard laboratory concentrations (0.5-10 uM in vitro; 0.1-1 mg/kg in vivo). In vitro cytotoxicity studies indicate no significant reduction in cell viability at concentrations up to 10 uM for 24 hours. In vivo, no adverse effects or mortality are reported at standard tracing doses. However, as an iodide salt and a lipophilic cation, high concentrations may cause mitochondrial membrane potential dissipation and cytotoxicity. The compound is for research use only and not intended for clinical applications. Standard laboratory precautions, including the use of gloves and eye protection, should be followed when handling this dye.
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| References | |
| Additional Infomation |
4-Di-10-ASP is a research-grade fluorescent dye with no approved clinical applications. Its excitation maximum is 485 nm and emission maximum is 620 nm, producing orange to red fluorescence. The dye is provided as an orange to red solid powder and should be stored at 4degC in a sealed container, protected from moisture and light. Stock solutions in DMSO or ethanol should be stored at -20degC for up to 6 months. The dye is soluble in organic solvents such as DMSO, DMF, and ethanol, but has limited aqueous solubility. It is commonly used in combination with other fluorescent probes for multiplexed imaging of cellular membranes and organelles. Synonyms include N,N-didecyl-4-[(E)-2-(1-methylpyridin-1-ium-4-yl)ethenyl]aniline iodide.
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| Molecular Formula |
C34H55N2+.I-
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|---|---|
| Molecular Weight |
618.7179
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| Exact Mass |
618.341
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| CAS # |
95378-73-7
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| PubChem CID |
5706739
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| Appearance |
Orange to red solid powder
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| LogP |
6.773
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
21
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| Heavy Atom Count |
37
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| Complexity |
479
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCCCCCCCCCN(CCCCCCCCCC)C1=CC=C(C=C1)/C=C/C2=CC=[N+](C=C2)C.[I-]
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| InChi Key |
PPAYPDQCRKDOKR-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C34H55N2.HI/c1-4-6-8-10-12-14-16-18-28-36(29-19-17-15-13-11-9-7-5-2)34-24-22-32(23-25-34)20-21-33-26-30-35(3)31-27-33;/h20-27,30-31H,4-19,28-29H2,1-3H3;1H/q+1;/p-1
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| Chemical Name |
N,N-didecyl-4-[(E)-2-(1-methylpyridin-1-ium-4-yl)ethenyl]aniline;iodide
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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: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 : ~33.33 mg/mL (~53.87 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 | 1.6162 mL | 8.0812 mL | 16.1624 mL | |
| 5 mM | 0.3232 mL | 1.6162 mL | 3.2325 mL | |
| 10 mM | 0.1616 mL | 0.8081 mL | 1.6162 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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