FCCP
(Synonyms: 碳酰氰-4-三氟甲氧基苯腙,Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone) 目录号 : GC14328
FCCP是一种质子载体（H+离子载体），也是一种强磷酸化解偶联剂，促进质膜和线粒体膜的去极化。
Cas No.:370-86-5
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
HEK 293 cells |
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Preparation Method |
Preceding the addition of drugs, cultures were washed once with warm phosphate-buffered saline and then exposed to various treatments in Dulbecco s modified Eagle s medium supplemented with sodium pyruvate (1 mM). Cultures were then exposed to control medium, vehicle, or varying concentrations of FCCP (5 µM, 500 nM, or 50 nM in 0.05% ethanol) or baf A1 (1 µM in dimethyl sulfoxide). Four hours after addition of drugs, the medium was removed. |
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Reaction Conditions |
FCCP (5 µM, 500 nM, or 50 nM in 0.05% ethanol) for 4 hours |
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Applications |
FCCP inhibits APP catabolism but not maturation |
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References: [1]. Connop BP, Thies RL, et,al. Novel effects of FCCP [carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone] on amyloid precursor protein processing. J Neurochem. 1999 Apr;72(4):1457-65. doi: 10.1046/j.1471-4159.1999.721457.x. PMID: 10098849. |
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FCCP, a proton carrier (H+ ionophore), is also a powerful phosphoric acid defusing coupling agent, which promotes the depolarization of plasma membrane and mitochondrial membrane.FCCP can affect various activities in which cellular calcium ions participate, inhibit K+ background current and induce small inward current, decrease 0.1 unit pH, and induce intracellular Na+ to increase. FCCP can stimulate Mg2+-ATPase activity, inhibit β-amyloid formation, and simulate the physiological effect of glutamate receptor agonist NMDA on mitochondrial superoxide.
Exposure of K695sw cells to the protonophore FCCP resulted in a concentration-dependent decrease in both Aβ release and the formation of the cell-associated C99 fragment. Production of the ectodomain fragment APPsα was only affected by exposure to the highest concentration of FCCP, whereas neither total cellular levels of APP nor the maturation of APP via N /O -linked glycosylation appeared to be affected by any concentration of FCCP used[1]. Pre-exposure to 200 nM FCCP during 120 min protects and enhances the follicle integrity in cat ovarian tissue during short-term in vitro culture[2]. Concentrations of FCCP that cause mitochondrial oxidation without depolarisation are cardioprotective. Higher FCCP concentrations dissipate mitochondrial membrane potential and exacerbate injury[3]. FCCP activated ionic currents and depolarized the plasma membrane potential in a dose-dependent manner. Neither the removal of extracellular Ca2+ nor pretreatment with BAPTA/AM affected the FCCP-induced currents, implying that the currents are not associated with the FCCP-induced intracellular [Ca2+]i increase[4]. Application of FCCP evoked a gradual increase in cell body [Ca2+]i that reached a level approximately 3-fold higher than baseline after 60 min. Moreover, FCCP released Ca2+ even when added after mitochondrial stores of Ca2+ had previously been emptied by an alternate method. FCCP, in addition to its recognized effect on mitochondrial Ca2+ sequestration, also releases Ca2+ from a non-mitochondrial store and is, therefore, unsuitable for use in an intact neuron to selectively inactivate mitochondrial Ca2+ uptake[5]. A further analysis of this effect on BHK21 cells has shown that a decrease in the number of microtubules can be observed 15 min after adding FCCP and there is complete disruption after 60 min. Regrowth of microtubules was initiated 30 min after removal of FCCP, in marked contrast with the rapid reversion observed when microtubules are disrupted by nocodazole[6].
References:
[1]. Connop BP, Thies RL, et,al. Novel effects of FCCP [carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone] on amyloid precursor protein processing. J Neurochem. 1999 Apr;72(4):1457-65. doi: 10.1046/j.1471-4159.1999.721457.x. PMID: 10098849.
[2]. Tanpradit N, Chatdarong K, et,al. Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) pre-exposure ensures follicle integrity during in vitro culture of ovarian tissue but not during cryopreservation in the domestic cat model. J Assist Reprod Genet. 2016 Dec;33(12):1621-1631. doi: 10.1007/s10815-016-0810-5. Epub 2016 Sep 17. PMID: 27639998; PMCID: PMC5171893.
[3]. Brennan JP, Berry RG, et,al. FCCP is cardioprotective at concentrations that cause mitochondrial oxidation without detectable depolarisation. Cardiovasc Res. 2006 Nov 1;72(2):322-30. doi: 10.1016/j.cardiores.2006.08.006. Epub 2006 Aug 16. PMID: 16979603.
[4]. Park KS, Jo I, et,al. FCCP depolarizes plasma membrane potential by activating proton and Na+ currents in bovine aortic endothelial cells. Pflugers Arch. 2002 Jan;443(3):344-52. doi: 10.1007/s004240100703. Epub 2001 Oct 6. PMID: 11810202.
[5]. Jensen JR, Rehder V. FCCP releases Ca2+ from a non-mitochondrial store in an identified Helisoma neuron. Brain Res. 1991 Jun 14;551(1-2):311-4. doi: 10.1016/0006-8993(91)90947-t. PMID: 1913158.
[6]. Maro B, Marty MC, et,al. In vivo and in vitro effects of the mitochondrial uncoupler FCCP on microtubules. EMBO J. 1982;1(11):1347-52. doi: 10.1002/j.1460-2075.1982.tb01321.x. PMID: 6765194; PMCID: PMC553215.
FCCP是一种质子载体（H+离子载体），也是一种强磷酸化解偶联剂，促进质膜和线粒体膜的去极化。FCCP可以影响细胞钙离子参与的各种活动，抑制K+背景电流并诱导小的内向电流，降低0.1个单位的pH值，诱导细胞内Na+增加。 FCCP可刺激Mg2+-ATP酶活性，抑制β-淀粉样蛋白形成，模拟谷氨酸受体激动剂NMDA对线粒体超氧化物的生理作用。
将 K695sw 细胞暴露于质子载体 FCCP 会导致 Aβ 释放和细胞相关 C99 片段形成的浓度依赖性降低。胞外域片段 APPsα 的产生仅受暴露于最高浓度 FCCP 的影响，而 APP 的总细胞水平和通过 N /O 连接糖基化的 APP 成熟似乎不受所用任何浓度 FCCP 的影响 [1]。在 120 分钟内预暴露于 200 nM FCCP 可在短期体外培养过程中保护和增强猫卵巢组织中的卵泡完整性[2]。引起线粒体氧化而不去极化的 FCCP 浓度具有心脏保护作用。较高的 FCCP 浓度会耗散线粒体膜电位并加剧损伤[3]。 FCCP 激活离子电流并以剂量依赖性方式去极化质膜电位。细胞外 Ca2+ 的去除和 BAPTA/AM 的预处理都不影响 FCCP 诱导的电流，这意味着电流与 FCCP 诱导的细胞内 [Ca2+]i 增加无关[4]。 FCCP 的应用引起细胞体 [Ca2+]i 的逐渐增加，在 60 分钟后达到比基线高约 3 倍的水平。此外，即使在先前通过替代方法清空 Ca2+ 的线粒体储存后添加，FCCP 也会释放 Ca2+。 FCCP 除了公认的对线粒体 Ca2+ 螯合的作用外，还从非线粒体储存中释放 Ca2+，因此不适合在完整神经元中用于选择性灭活线粒体 Ca2+ 摄取[5] .进一步分析这种对 BHK21 细胞的影响表明，在添加 FCCP 后 15 分钟可以观察到微管数量减少，60 分钟后完全破坏。微管在去除 FCCP 后 30 分钟开始再生，这与诺考达唑破坏微管时观察到的快速恢复形成鲜明对比[6]。
| Cas No. | 370-86-5 | SDF | |
| 别名 | 碳酰氰-4-三氟甲氧基苯腙,Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone | ||
| 化学名 | (4-(trifluoromethoxy)phenyl)carbonohydrazonoyl dicyanide | ||
| Canonical SMILES | FC(F)(OC1=CC=C(C=C1)N/N=C(C#N)/C#N)F | ||
| 分子式 | C10H5F3N4O | 分子量 | 254.17 |
| 溶解度 | ≥ 56.6 mg/mL in DMSO with ultrasonic, ≥ 25 mg/mL in EtOH with ultrasonic | 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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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.9344 mL | 19.6719 mL | 39.3437 mL |
| 5 mM | 0.7869 mL | 3.9344 mL | 7.8687 mL |
| 10 mM | 0.3934 mL | 1.9672 mL | 3.9344 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 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 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
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，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
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。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Protonophore FCCP provides fitness advantage to PDR-deficient yeast cells
J Bioenerg Biomembr2020 Oct;52(5):383-395.PMID: 32808242DOI: 10.1007/s10863-020-09849-1
Pleiotropic drug resistance (PDR) plasma membrane transporters mediate xenobiotic efflux from the cells and thereby help pathogenic microorganisms to withstand antimicrobial therapies. Given that xenobiotic efflux is an energy-consuming process, cells with upregulated PDR can be sensitive to perturbations in cellular energetics. Protonophores dissipate proton gradient across the cellular membranes and thus increase ATP spendings to their maintenance. We hypothesised that chronic exposure of yeast cells to the protonophores can favour the selection of cells with inactive PDR. To test this, we measured growth rates of the wild type Saccharomyces cerevisiae and PDR-deficient Δpdr1Δpdr3 strains in the presence of protonophores carbonyl cyanide-p-trifluoromethoxyphenylhydrazone(FCCP),pentachlorophenol (PCP) and niclosamide (NCA). Although the protonophore-induced respiration rates of these two strains were similar, the PDR-deficient strain outperformed the control one in the growth rate on non-fermentable carbon source supplemented with low concentrations of FCCP. Thus, active PDR can be deleterious under conditions of partially uncoupled oxidative-phosphorylation. Furthermore, our results suggest that tested anionic protonophores are poor substrates of PDR-transporters. At the same time, protonophores imparted azole tolerance to yeasts, pointing that they are potent PDR inducers. Interestingly, protonophore PCP led to a persistent increase in the levels of a major ABC-transporter Pdr5p, while azole clotrimazole induced only a temporary increase. Together, our data provides an insight into the effects of the protonophores in the eukaryotes at the cellular level and support the idea that cells with activated PDR can be selected out upon conditions of energy limitations.
Current mechanistic insights into the CCCP-induced cell survival response
Biochem Pharmacol2018 Feb;148:100-110.PMID: 29277693DOI: 10.1016/j.bcp.2017.12.018
The ring-substituted derivatives of carbonyl cyanide phenylhydrazone, CCCP and FCCP, are routinely used for the analysis of the mitochondrial function in living cells, tissues, and isolated mitochondrial preparations. CCCP and FCCP are now being increasingly used for investigating the mechanisms of autophagy by inducing mitochondrial degradation through the disruption of the mitochondrial membrane potential (ΔΨm). Sustained perturbation of ΔΨm, which is normally tightly controlled to ensure cell proliferation and survival, triggers various stress pathways as part of the cellular adaptive response, the main components of which are mitophagy and autophagy. We here review current mechanistic insights into the induction of mitophagy and autophagy by CCCP and FCCP. In particular, we analyze the cellular modifications produced by the activation of two major pathways involving the signaling of the nuclear factor erythroid 2-related factor 2 (Nrf2) and the transcription factor EB (TFEB), and discuss the contribution of these pathways to the integrated cellular stress response.
'Mild Uncoupling' does not decrease mitochondrial superoxide levels in cultured cerebellar granule neurons but decreases spare respiratory capacity and increases toxicity to glutamate and oxidative stress
J Neurochem2007 Jun;101(6):1619-31.PMID: 17437552DOI: 10.1111/j.1471-4159.2007.04516.x
Cultured rat cerebellar granule neurons were incubated with low nanomolar concentrations of the protonophore carbonylcyanide-p-trifluoromethoxyphenyl hydrazone (FCCP) to test the hypothesis that 'mild uncoupling' could be neuroprotective by decreasing oxidative stress. To quantify the uncoupling, respiration and mitochondrial membrane potential (Deltapsi(m)) were determined in parallel as a function of FCCP concentration. Deltapsi(m) dropped by less than 10 mV before respiratory control was lost. Conditions for the valid estimation of matrix superoxide levels were determined from the rate of oxidation of the matrix-targeted fluorescent probe MitoSOX. No significant change in the level of matrix superoxide could be detected on addition of FCCP while respiratory control was retained, although cytoplasmic superoxide levels measured by dihydroethidium oxidation increased. 'Mild uncoupling' by 30 nmol/L FCCP did not alleviate neuronal dysregulation induced by glutathione depletion and significantly enhanced that due to menadione-induced oxidative stress. Low protonophore concentrations enhanced N-methyl-d-aspartate receptor-induced delayed calcium deregulation consistent with a decrease in the spare respiratory capacity available to match the bioenergetic demand of chronic receptor activation. It is concluded that the 'mild uncoupling' hypothesis is not supported by this model.
Mitochondrial uncouplers induce proton leak by activating AAC and UCP1
Nature2022 Jun;606(7912):180-187.PMID: 35614225DOI: 10.1038/s41586-022-04747-5
Mitochondria generate heat due to H+ leak (IH) across their inner membrane1. IH results from the action of long-chain fatty acids on uncoupling protein 1 (UCP1) in brown fat2-6 and ADP/ATP carrier (AAC) in other tissues1,7-9, but the underlying mechanism is poorly understood. As evidence of pharmacological activators of IH through UCP1 and AAC is lacking, IH is induced by protonophores such as 2,4-dinitrophenol (DNP) and cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP)10,11. Although protonophores show potential in combating obesity, diabetes and fatty liver in animal models12-14, their clinical potential for treating human disease is limited due to indiscriminately increasing H+ conductance across all biological membranes10,11 and adverse side effects15. Here we report the direct measurement of IH induced by DNP, FCCP and other common protonophores and find that it is dependent on AAC and UCP1. Using molecular structures of AAC, we perform a computational analysis to determine the binding sites for protonophores and long-chain fatty acids, and find that they overlap with the putative ADP/ATP-binding site. We also develop a mathematical model that proposes a mechanism of uncoupler-dependent IH through AAC. Thus, common protonophoric uncouplers are synthetic activators of IH through AAC and UCP1, paving the way for the development of new and more specific activators of these two central mediators of mitochondrial bioenergetics.