| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
Voltage-sensitive fluorescent dye
DiBAC4(3) targets the cellular membrane potential. It is a negatively charged, hydrophobic molecule that partitions into the lipid bilayer of cell membranes. In response to changes in membrane potential, the dye redistributes across the membrane. At resting (more negative) membrane potentials, the dye is excluded from the cell. Upon depolarization (less negative), the dye enters the cell and binds to intracellular proteins and lipids, resulting in an increase in fluorescence intensity. This property makes it a valuable tool for monitoring membrane potential changes in various cell types. |
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| ln Vitro |
Membrane hyperpolarization mediated by 10 μM Evans Blue (EB) in HEKBKα was clearly seen using DiBAC4(3), however the addition of 3 mM tetraethylammonium chloride (TEA) led in a significantly lesser change in membrane potential (MP). Electrodes measure slowly. The peak periods of hyperpolarization assessed by microelectrodes and DiBAC4(3) were 2.3±0.9 s (n=4) and 35.0±2.6 s (n=12, P<0.01) respectively [1].
In vitro, DiBAC4(3) is widely used to measure changes in cellular membrane potential in real-time. It is used in high-throughput screening to identify modulators of ion channels and transporters. The dye's fluorescence is excited at 490 nm and emits at 505 nm. Its response is slow, making it suitable for measuring sustained changes in membrane potential rather than rapid action potentials. It is also used to monitor changes in the plasma membrane potential of various cell types, including neurons, muscle cells, and epithelial cells. |
| ln Vivo |
In vivo, DiBAC4(3) is not used as a therapeutic agent but as a research tool. Its application is primarily in ex vivo or in vitro settings to study cellular physiology. It could potentially be used in vivo to monitor membrane potential in specific tissues, but this is not a common application. Its primary role is as a laboratory reagent for fluorescence-based assays.
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| Enzyme Assay |
In vitro non-cell assays for DiBAC4(3) are not enzyme-based. The dye's properties are characterized by its spectral characteristics (excitation and emission maxima) and its response to changes in membrane potential in artificial membrane systems, such as liposomes or lipid bilayers. Its purity is assessed by HPLC.
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| Cell Assay |
Before the fluorescence measurements, cells were incubated in KRH (Krebs-Ringer-HEPES) buffer containing with 100 nM DiBAC4(3) for 20 min at room temperature. The stained cells were used for experiments without washing. The fluorescence emission was collected using a 505 nm dicroic mirror and a BA filter (>520 nm)[1].
In vitro cell-based assays for DiBAC4(3) are the primary application of this compound. Cells are loaded with the dye, and changes in fluorescence intensity are measured using a fluorescence plate reader, flow cytometer, or fluorescence microscope in response to various stimuli. The dye is used to monitor the effects of drugs, toxins, or other treatments on membrane potential. It is a key tool in ion channel drug discovery. |
| Animal Protocol |
In vivo animal studies are not typically performed with DiBAC4(3), as it is a fluorescent dye used in cell-based assays. Its use is limited to in vitro and ex vivo applications, such as imaging of tissue slices.
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| ADME/Pharmacokinetics |
DiBAC4(3) has a molecular weight of 516.63 g/mol and a molecular formula of C₂₇H₄₀N₄O₆. It is an orange to red powder to crystal. It is soluble in DMSO or DMF. It should be stored at -20°C and protected from light. Its stability in solution is limited, and it should be prepared fresh before use.
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| Toxicity/Toxicokinetics |
DiBAC4(3) is considered a hazardous chemical and should be handled with appropriate safety precautions. It is an irritant and should be used in a well-ventilated area. It is not intended for human or veterinary use.
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| References | |
| Additional Infomation |
This study evaluated the application of the voltage-sensitive fluorescent dye bis-(1,3-dibutylbarbituric acid)-trimethoxyxanthol (DiBAC4(3)) in membrane potential (MP) measurement in HEK293 cells. HEK293 cells express the α subunit or α+β1 subunit of a high-conductivity calcium-activated potassium (BK) channel (denoted as HEKBKα and HEKBKαβ, respectively). Under voltage-clamped conditions, the fluorescence intensity of DiBAC4(3) was measured at different membrane potentials to calibrate and semi-quantitatively estimate the absolute membrane potential. The resting membrane potential measured by DiBAC4(3) was roughly equivalent to the membrane potential recorded by the microelectrode; the membrane potential of HEKBKα cells was 10-20 mV lower than that of native HEK cells. In HEKBKα cells, 10 μM Evans blue (a BK channel opener)-induced membrane hyperpolarization was detectable by DiBAC4(3). NS-1619 is another BK channel opener that induces a slow but significant change in F/F(K) even in native HEK cells, while simultaneously detecting the opening effect of BK channels. In HEKBKαβ cells, oscillatory membrane hyperpolarization can be induced by increasing intracellular Ca2+ concentration and applying 10 μM acetylcholine. However, this oscillatory hyperpolarization can only be detected by DiBAC4(3) and appears as a slow hyperpolarization. It can be concluded that the relatively slow effect of BK channel regulators can be semi-quantitatively measured using DiBAC4(3) in HEKBK cells, but its limited temporal resolution and potential artifacts should be taken into account. [1]
DiBAC4(3) is a voltage-sensitive fluorescent dye used to measure changes in cellular membrane potential. It is a sensitive slow-response probe. It is widely used in high-throughput screening and cell-based assays. It is not approved for clinical use and is intended for research purposes only. |
| Molecular Formula |
C27H40N4O6
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|---|---|
| Molecular Weight |
516.629707336426
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| Exact Mass |
516.29
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| Elemental Analysis |
C, 62.77; H, 7.80; N, 10.84; O, 18.58
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| CAS # |
70363-83-6
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| Appearance |
Pink to red solid powder
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| LogP |
2.901
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| SMILES |
C1(=O)N(CCCC)C(=O)/C(=C\C=CC2C(=O)N(CCCC)C(=O)N(CCCC)C2=O)/C(=O)N1CCCC
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| InChi Key |
ATJCYSYHWGQAET-BUHFOSPRSA-N
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| InChi Code |
InChI=1S/C27H40N4O6/c1-5-9-16-28-22(32)20(23(33)29(26(28)36)17-10-6-2)14-13-15-21-24(34)30(18-11-7-3)27(37)31(25(21)35)19-12-8-4/h13-15,20H,5-12,16-19H2,1-4H3/b14-13+
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| Chemical Name |
(E)-1,3-dibutyl-5-(3-(1,3-dibutyl-2,4,6-trioxohexahydropyrimidin-5-yl)allylidene)pyrimidine-2,4,6(1H,3H,5H)-trione
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| Synonyms |
DiBAC4(3); DiBAC4-(3); DiBAC4 (3);
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : ~50 mg/mL (~96.78 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 | 1.9356 mL | 9.6781 mL | 19.3562 mL | |
| 5 mM | 0.3871 mL | 1.9356 mL | 3.8712 mL | |
| 10 mM | 0.1936 mL | 0.9678 mL | 1.9356 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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