DiBAC4(3) (Synonyms: Bis(1,3-Dibutylbarbituric Acid) Trimethine Oxonol) |
Catalog No.GC30140
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DiBAC4(3) is a negatively charged membrane potential-sensitive bis-oxonol dye with excitation maxima at approximately 490nm and its fluorescence emission is collected using a 520nm filter.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 70363-83-6
Sample solution is provided at 25 µL, 10mM.
DiBAC4(3) is a negatively charged membrane potential-sensitive bis-oxonol dye with excitation maxima at approximately 490nm and its fluorescence emission is collected using a 520nm filter[1]. DiBAC4(3) is a versatie tool in membrane potential monitoring, viability assays and antibiotic susceptibility testing.
In vitro, DiBAC4(3) was used to measure membrane potentials in HEK293 cells expressing recombinant large conductance Ca2+-activated K+ (BK) channels (HEKBKα and HEKBKβ ). Cells were incubated in KRH (Krebs-RingerHEPES) buffer containing with 100nM DiBAC4(3) for 20min at room temperature. The oscillatory membrane hyperpolarization induced in HEKBKαβ by was detected only as a slow hyperpolarization with DiBAC4(3)[2]. B. subtilis cells incubated with membrane-targeting antimicrobials were stained with 10μM DiBAC4(3) for 17h and observed under phase contrast and fluorescence microscopy to evaluate the effects of antimicrobial compounds on membrane integrity and function[3]. DiBAC4(3) (1μM) was used to assess cell viability and membrane potential in starved cultures of E. coli and Salmonella in seawater. Cells may maintain some membrane potential, they do not necessarily exhibit active respiration, especially in starved conditions[4].
References:
[1] Epps D E, Wolfe M L, Groppi V. Characterization of the steady-state and dynamic fluorescence properties of the potential-sensitive dye bis-(1,3-dibutylbarbituric acid)trimethine oxonol (Dibac4(3)) in model systems and cells. Chem Phys Lipids. 1994 Feb;69(2):137-50.
[2] Yamada A, Gaja N, Ohya S,et al. Usefulness and limitation of DiBAC4(3), a voltage-sensitive fluorescent dye, for the measurement of membrane potentials regulated by recombinant large conductance Ca2+-activated K+ channels in HEK293 cells. Jpn J Pharmacol. 2001 Jul;86(3):342-50.
[3] Winkel J D T, Gray D A, Seistrup K H. Analysis of Antimicrobial-Triggered Membrane Depolarization Using Voltage Sensitive Dyes. Front Cell Dev Biol. 2016 Apr 13:4:29.
[4] Amorós R L, Castel S, Riu J C, Rego J V. Assessment of E. coli and Salmonella viability and starvation by confocal laser microscopy and flow cytometry using rhodamine 123, DiBAC4(3), propidium iodide, and CTC. Cytometry. 1997 Dec 1;29(4):298-305.
The use of DiBAC4(3) in HEK 293T cells to measure membrane potentials：
1. Prepare the standard external solution, KRH (Krebs-RingerHEPES) solution having the following composition was used as the external solution: 127mM NaCl, 5mM KCl, 2mM CaCl2, 1.2mM MgCl2, 6mM glucose and 25mM HEPES. The pH was adjusted to 7.4 with NaOH.
2. Human embryonic kidney cell lines HEK293 cells transfected with BKαβ1 stably expressed functional BK channels (HEKBKαβ) .
3. Dissolve DiBAC4(3) to the KRH buffer to achieve a final concentration of 100nM.
4. Incubated cells in KRH buffer containing with 100nM DiBAC(3) for 20min at room temperature. The stained cells were used for experiments without washing.
5. Excite DiBAC4(3) at approximately 490nm and collect the emitted fluorescence using a 505nm dichroic mirror and a 520nm bandpass filter.
6. Use a Ca2+ imaging system (ARGUS- HiSCA) for data collection and analysis. The sampling interval of DiBAC(3) fluorescence measurements was in the range of 4 and 5s.
This protocol only provides a guideline, and should be modified according to your specific needs.
[1] Yamada A, Gaja N, Ohya S,et al. Usefulness and limitation of DiBAC4(3), a voltage-sensitive fluorescent dye, for the measurement of membrane potentials regulated by recombinant large conductance Ca2+-activated K+ channels in HEK293 cells. Jpn J Pharmacol. 2001 Jul;86(3):342-50.
| Cas No. | 70363-83-6 | SDF | |
| Synonyms | Bis(1,3-Dibutylbarbituric Acid) Trimethine Oxonol | ||
| Canonical SMILES | O=C1N(CCCC)C(/C(C(N1CCCC)=O)=C/C=C/C(C(N2CCCC)=O)C(N(CCCC)C2=O)=O)=O | ||
| Formula | C27H40N4O6 | M.Wt | 516.63 |
| Solubility | DMSO : ≥ 60 mg/mL (116.14 mM) | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.9356 mL | 9.6781 mL | 19.3562 mL |
| 5 mM | 387.1 μL | 1.9356 mL | 3.8712 mL |
| 10 mM | 193.6 μL | 967.8 μL | 1.9356 mL |
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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
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- Purity: >97.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 12 reference(s) in Google Scholar.)