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CCCP 目录号 GC14727

uncoupler of oxidative phosphorylation

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Sample solution is provided at 25 µL, 10mM.

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实验参考方法

Cell experiment:

MEFs (5×105), Raw264.7 cells (1×106), and HeLa cells stable expressing STING (1.5×105) are stimulated with DMXAA (100 μg/mL) for 2 or 3 h, or transfected with c-di-GMP (5 μM), cGAMP (5 μg/mL), or poly (dA:dT) (2 μg/mL) for 6 h. CCCP (50 μM) is co-treated with DMXAA (100 μg/mL), or treated for the last 5 h in case of treatment of c-di-GMP or poly (dA:dT)[1].

Animal experiment:

Mice[2] Female Balb/c mice n=6, per dosing group weighing 20-25 g are rendered neutropenic with 2 intraperitoneal injections of cyclophosphamide 150 mg/kg.bw and 100 mg/kg.bw on 4 days and 1 day prior to bacterial infection. 0.1 mL of the 106 CFU/mL bacterial suspension is injected into right posterior thigh muscle. After 2 h post-infection mice are treated with PPEF (3 mg/kg.bw), CCCP (3 mg/kg.bw) and in combination PPEF+CCCP (3 mg/kg.bw+3 mg/kg.bw) dissolved in 0.1 mL sterile water by single bolus intravenous injection. Twenty-four hours after antibacterial administration, the mice are humanely sacrificed. Right thigh muscles from each mouse are aseptically collected, homogenized and serially diluted and processed for quantitative cultures. Rats[3] Rats are randomly divided into three groups. One group is euthanized 15 min after a dose of 12.5 MBq (337.8 μCi) 99mTc-MIBI injection (n=6). The other two groups are administered 4 mg/kg CCCP (CCCP group; n=7) or vehicle (vehicle group; n=7) by intraperitoneal (i.p.) injection 90 min after the same dose of 99mTc-MIBI injection and are euthanized after an additional 90 min (180 min after the 99mTc-MIBI injection). Hearts are excised and weighed, and radioactivity is measured between 110 and 170 keV with an auto-well gamma counter. 99mTc-MIBI signals are corrected for physical decay (half-life=6 h).

References:

[1]. Kwon D, et al. Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) suppresses STING-mediated DNA sensing pathway through inducing mitochondrial fission. Biochem Biophys Res Commun. 2017 Aug 30. pii: S0006-291X(17)31704-7.
[2]. Sinha D, et al. Synergistic efficacy of Bisbenzimidazole and Carbonyl Cyanide 3-Chlorophenylhydrazonecombination against MDR bacterial strains. Sci Rep. 2017 Mar 17;7:44419.
[3]. Kawamoto A, et al. Measurement of technetium-99m sestamibi signals in rats administered a mitochondrial uncoupler and in a rat model of heart failure. PLoS One. 2015 Jan 16;10(1):e0117091.
[4]. Kondapalli C, et al. PINK1 is activated by mitochondrial membrane potential depolarization and stimulates Parkin E3 ligase activity by phosphorylating Serine 65. Open Biol. 2012 May;2(5):120080.

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Chemical Properties

Cas No. 555-60-2 SDF
别名 N/A
化学名 (3-chlorophenyl)carbonohydrazonoyl dicyanide
Canonical SMILES ClC1=CC(N/N=C(C#N)/C#N)=CC=C1
分子式 C9H5ClN4 分子量 204.62
溶解度 ≥20.5mg/mL in DMSO 储存条件 Store at RT
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request

产品描述

IC50: N/A

CCCP (carbonyl cyanide m-chlorophenyl hydrazine) is widely used uncoupler of oxidative phosphorylation.

The uncoupler of oxidative phosphorylation blocks ATP synthesis by collapsing the proton motive force.

In vitro: A genetic β-galactoside reporter system with a disk diffusion assay on MacConkey Lactose agar petri plates to monitor maintenance of the bacteriophage λ prophage state and viral induction in Escherichia coli K-12. It presented evidence that the phage λ major lytic promoters, pL and pR, are activated via cells containing the reporters following exposure to the energy poison CCCP. Requirement of expression of the λ lytic promoters in response to CCCP is host RecA function and an auto-cleavable CI repressor, as does SOS induction of the λ prophage occurring by a DNA damage-dependent pathway. CCCP-mediated activation of the λ lytic promoters required λ Cro function. CCCP does not modulate an sfi-lacZ SOS reporter [1]. The uncoupler CCCP blocks oxidative phosphorylation by damaging the proton gradient. An anion CCCP can bind a proton. but CCCP with a delocalized negative charge allows it to cross the lipid bilayer in the unprotonated form, while weak acids cross the hydrophobic membrane only when protonated. Many protons can be transported by one molecule of CCCP across the inner membrane. It is described that a genetic reporter system to monitor maintenance of the λ prophage state and viral induction, and evidence is presented that it activates the major leftward and rightward lytic promoters of the λ prophage that cells are exposed to the energy poison CCCP.

In vivo: So far, no study in vivo has been conducted.

Clinical trial: So far, no clinical study has been conducted.

Reference:
[1].  Thomason LC, Court DL. Evidence that bacteriophage λ lysogens may induce in response to the proton motive force uncoupler CCCP. FEMS Microbiol Lett. 2016 Feb;363(3).