DiBAC4(3)
(Synonyms: 双(1,3-二巴比妥酸)-三次甲基氧烯洛尔) 目录号 : GC30140
A fluorescent membrane potential indicator
Cas No.:70363-83-6
Sample solution is provided at 25 µL, 10mM.
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- Purity: >98.00%
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Cell experiment: | Prior to the fluorescence measurements, cells are incubated in KRH (Krebs-Ringer-HEPES) buffer containing with 100 nM DiBAC4(3) for 20 min at room temperature. The stained cells are used for experiments without washing. The fluorescence emission is collected using a 505 nm dicroic mirror and a BA filter (>520 nm)[1]. |
References: [1]. Yamada A, 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. | |
DiBAC4(3) is a negatively charged fluorescent membrane potential indicator.1 Upon membrane depolarization, DiBAC4(3) enters the cytosol, binds to lipid membranes and intracellular proteins, and displays excitation/emission maxima of 490/516 nm, respectively.1,2 It has been used to measure the membrane potential of live or fixed mammalian and bacterial cells by flow cytometry or fluorescence microscopy.2,3,4
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 cellsJpn. J. Pharmacol.86(3)342-350(2001) 2.Warren, E.A.K., and Payne, C.K.Cellular binding of nanoparticles disrupts the membrane potentialRSC Adv.5(18)13660-13666(2015) 3.Klapperstück, T., Glanz, D., Klapperstück, M., et al.Methodological aspects of measuring absolute values of membrane potential in human cells by flow cytometryCytometry A.75(7)593-608(2009) 4.Mason, D.J., Allman, R., Stark, J.M., et al.Rapid estimation of bacterial antibiotic susceptibility with flow cytometryJ. Microsc.176(Pt 1)8-16(1994)
| Cas No. | 70363-83-6 | SDF | |
| 别名 | 双(1,3-二巴比妥酸)-三次甲基氧烯洛尔 | ||
| Canonical SMILES | O=C1N(CCCC)C(/C(C(N1CCCC)=O)=C/C=C/C(C(N2CCCC)=O)C(N(CCCC)C2=O)=O)=O | ||
| 分子式 | C27H40N4O6 | 分子量 | 516.63 |
| 溶解度 | DMSO : ≥ 60 mg/mL (116.14 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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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 |
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| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
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1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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DiBAC₿3) hits a "sweet spot" for the activation of arterial large-conductance Ca²ⁿactivated potassium channels independently of the β₿subunit
Am J Physiol Heart Circ Physiol 2013 Jun 1;304(11):H1471-82.23542916 PMC3680725
The voltage-sensitive dye bis-(1,3-dibutylbarbituric acid)trimethine oxonol [DiBAC₿3)] has been reported as a novel large-conductance Ca²ⁿactivated Kⁿ(BK) channel activator with selectivity for its β₿ or β₿subunits. In arterial smooth muscle, BK channels are formed by a pore-forming α-subunit and a smooth muscle-abundant regulatory β₿subunit. This tissue specificity has driven extensive pharmacological research aimed at regulating arterial tone. Using animals with a disruption of the gene for the β₿subunit, we explored the effects of DiBAC₿3) in native channels from arterial smooth muscle. We tested the hypothesis that, in native BK channels, activation by DiBAC₿3) relies mostly on its α-subunit. We studied BK channels from wild-type and transgenic β₿knockout mice in excised patches. BK channels from brain arteries, with or without the β₿subunit, were similarly activated by DiBAC₿3). In addition, we found that saturating concentrations of DiBAC₿3) (~30 μM) promote an unprecedented persistent activation of the channel that negatively shifts its voltage dependence by as much as -300 mV. This "sweet spot" for persistent activation is independent of Ca²ⁿand/or the β₁₋₿subunits and is fully achieved when DiBAC₿3) is applied to the intracellular side of the channel. Arterial BK channel response to DiBAC₿3) varies across species and/or vascular beds. DiBAC₿3) unique effects can reveal details of BK channel gating mechanisms and help in the rational design of BK channel activators.
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):342-50.11488436 10.1254/jjp.86.342
The usefulness of bis-(1,3-dibutylbarbituric acid)-trimethine oxonol (DiBAC4(3)), a voltage-sensitive fluorescent dye, for the measurement of membrane potentials (MPs) was evaluated in HEK293 cells, where alpha or alpha plus beta1 subunits of large conductance Ca2+-activated K+ (BK) channels were expressed (HEKBK alpha and HEKBK alphabeta). The fluorescent intensity of DiBAC4(3) was measured at various potentials under voltage-clamp for calibration to estimate the absolute MP semi-quantitatively. The resting MPs measured with DiBAC4(3) were roughly comparable to those recorded with a microelectrode; the MP in HEKBK alphabeta was 10-20 mV more negative than that in native HEK. In HEKBK alpha, the membrane hyperpolarization induced by 10 microM Evans blue, a BK channel opener, was detected with DiBAC4(3). NS-1619, another BK channel opener, induced gradual but substantial change in F/F(K) even in native HEK, while the BK channel opening effect was detected. Oscillatory membrane hyperpolarization was induced in HEKBK alphabeta by application of 10 microM acetylcholine via increase in intracellular Ca2+ concentration. The oscillatory hyperpolarization was, however, detected only as a slow hyperpolarization with DiBAC4(3). It can be concluded that relatively slow effects of BK channel modulators can be semi-quantitatively measured by use of DiBAC4(3) in HEKBK, while the limited temporal resolution and possible artifacts should be taken into account.
The smooth muscle-type β1 subunit potentiates activation by DiBAC4(3) in recombinant BK channels
Channels (Austin) 2014;8(1):95-102.24299688 PMC4048348
Large-conductance Ca(2+)-activated (BK) channels, expressed in a variety of tissues, play a fundamental role in regulating and maintaining arterial tone. We recently demonstrated that the slow voltage indicator DiBAC4(3) does not depend, as initially proposed, on the β 1 or β 4 subunits to activate native arterial smooth muscle BK channels. Using recombinant mslo BK channels, we now show that the β 1 subunit is not essential to this activation but exerts a large potentiating effect. DiBAC4(3) promotes concentration-dependent activation of BK channels and slows deactivation kinetics, changes that are independent of Ca(2+). Kd values for BK channel activation by DiBAC4(3) in 0 mM Ca(2+) are approximately 20 μM (α) and 5 μM (α+β 1), and G-V curves shift up to -40 mV and -110 mV, respectively. β1 to β2 mutations R11A and C18E do not interfere with the potentiating effect of the subunit. Our findings should help refine the role of the β 1 subunit in cardiovascular pharmacology.
Live Imaging of Planarian Membrane Potential Using DiBAC4(3)
CSH Protoc 2008 Oct 1;2008:pdb.prot5055.21356693 10.1101/pdb.prot5055
INTRODUCTIONThis protocol describes how to use the anionic membrane voltage-reporting dye DiBAC(4)(3) to generate images of cell membrane potential in live planarians. These images qualitatively reveal variations in time-averaged membrane potential across different regions of the organism. Changes in these images due to experimental treatments reveal how the particular treatment affects this physiological parameter. This method is a great improvement over standard electrophysiological techniques, which cannot be used to gain an understanding of the electrical properties of an entire worm or a regenerating fragment, due to small cell size and large cell number. When the proper controls are performed, this technique is a very powerful and simple way to gather physiologic data.
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):137-50.8181103 10.1016/0009-3084(94)90035-3
The steady-state and dynamic fluorescence properties of the membrane potential-sensitive bis-oxonol dye DiBAC4(3) were characterized in vitro using model ligand systems and in vivo in A10 smooth muscle cells by fluorescence microscopy in conjunction with the ACAS imaging system. In the latter system the dye responds to potassium ion-induced jumps in membrane potential with changes in its fluorescence intensity, which follow pseudo-first-order kinetics. The relationship between the magnitude of the changes and the corresponding rate constants excludes the possibility that a simple, one-step equilibrium between extracellular and cytoplasmic dye would be sufficient to account for this phenomenon. The necessity of invoking an additional step suggested that the redistribution of the free dye between the cytoplasm and the exocellular medium is rapid and that the slow step associated with the fluorescence changes may be the interaction of the dye with proteins in the cytoplasm, along the lines proposed by Bräuner et al. (Biochim. Biophys. Acta 771 (1984), 208, 216). The interaction of the dye with BSA and with egg lecithin SUVs was studied as a model for the in vivo phenomenon. The dependence of fluorescence intensity changes on the concentrations of the reagents shows the formation of a reversible dye/albumin complex with a 2/1-stoichiometry, with Kd = 0.03 +/- 0.01 microM and a reversible adsorption to the SUVs with Kd 0.45 +/- 0.08 microM. The fluorescence lifetime of the dye in solution, < 100 ps, results in a high solution steady-state anisotropy. The latter decreases considerably upon binding to BSA, SUVs and A10 cells concomitant with a large increase in the lifetime. With such a short lifetime of the free dye, selective gating of the excitation source or the photodetector should eliminate the high background of the unbound dye and thereby enhance the sensitivity of the system.