GLPBio

CCCP

Catalog No.: GC14727Purity:>99.50%Synonyms: Carbonyl cyanide m-chlorophenyl hydrazone, NSC 88124
COASDSData Sheet
CCCP isa protonophore altering the permeability of mitochondria inner membrane to protons.

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CCCP
Cas No.: 555-60-2
100mgIn Stock
$44.00
1
500mgIn Stock
$136.00
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1gIn Stock
$200.00
1
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Citations

6 publicationsPublications from researchers using this product
IF 12

Description

CCCP is a protonophore altering the permeability of mitochondria inner membrane to protons[1]. CCCP increases membrane proton conductance by several orders of magnitude, thereby causing mitochondrial depolarization and uncoupling of respiration[2]. CCCP disrupts the cytosolic complex between the actin-binding protein Keap1 and the transcription factor Nrf2, releases Nrf2, and enables Nrf2 translocation to the nucleus, where Nrf2 activates the antioxidant response element (ARE)[3]. CCCP has been widely used in cell metabolism research to reduce ATP production and reverse bacterial resistance to antibiotics[4].

In vitro, CCCP treatment for 72 hours significantly inhibited the cell viability of 92-1, OMM1, and OMM2.3 cells, with the IC50 values being 6.5µM, 10.7µM, and 11.1µM, respectively[5]. Incubation with 25µM CCCP for 12 hours markedly decreased the phosphorylation of mTORC1 substrates eIF4E-BP1 and S6K in HepG2 cells and promoted autophagy as well as mitochondrial dysfunction[6]. Treatment with 100µM CCCP for 12 hours induced Keap1 degradation and apoptosis in p62-/- mouse embryonic fibroblast (MEF) cells, accompanied by an increase in ROS accumulation and the expression levels of cleaved PARP and caspase-3[7].

In vivo, CCCP treatment via intraperitoneal injection at a dose of 1mg/kg/day for 21 days improved the neurological deficits and motor dysfunction in the mouse model of intracerebral hemorrhage (ICH), and enhanced spatial learning and memory functions[8]. Intraperitoneal injection of CCCP at a dose of 1mg/kg/day for 2 weeks induced hepatic fibrosis without causing liver injury in mice[9].

References:

[1] Kasianowicz J, Benz R, McLaughlin S. The kinetic mechanism by which CCCP (carbonyl cyanide m-chlorophenylhydrazone) transports protons across membranes[J]. The Journal of membrane biology, 1984, 82(2): 179-190.

[2] Heytler P G. Uncoupling of oxidative phosphorylation by carbonyl cyanide phenylhydrazones. I. Some characteristics of m-CI-CCP action on mitochondria and chloroplasts[J]. Biochemistry, 1963, 2(2): 357-361.

[3] Kane M S, Paris A, Codron P, et al. Current mechanistic insights into the CCCP-induced cell survival response[J]. Biochemical pharmacology, 2018, 148: 100-110.

[4] Osei Sekyere J, Amoako D G. Carbonyl cyanide m-chlorophenylhydrazine (CCCP) reverses resistance to colistin, but not to carbapenems and tigecycline in multidrug-resistant Enterobacteriaceae[J]. Frontiers in microbiology, 2017, 8: 228.

[5] Xie M, Gu X, Zhao Z, et al. Artemisinin synergizes with CCCP in autophagic cell death induction via ER stress in uveal melanoma[J]. Iscience, 2025, 28(8).

[6] Koncha R R, Ramachandran G, Sepuri N B V, et al. CCCP‐induced mitochondrial dysfunction–characterization and analysis of integrated stress response to cellular signaling and homeostasis[J]. The FEBS journal, 2021, 288(19): 5737-5754.

[7] Park J S, Kang D H, Bae S H. p62 prevents carbonyl cyanide m-chlorophenyl hydrazine (CCCP)-induced apoptotic cell death by activating Nrf2[J]. Biochemical and biophysical research communications, 2015, 464(4): 1139-1144.

[8] Xiaoyu L, Dandan L, Tianzhao O, et al. Resolvin D1 combined with exercise rehabilitation alleviates neurological injury in mice with intracranial hemorrhage via the BDNF/TrkB/PI3K/AKT pathway[J]. Scientific Reports, 2024, 14(1): 31447.

[9] Lee J H, Seo K H, Yang J H, et al. CCCP induces hepatic stellate cell activation and liver fibrogenesis via mitochondrial and lysosomal dysfunction[J]. Free Radical Biology and Medicine, 2024, 225: 181-192.

Protocol

Cell experiment [1]:

Cell lines

 OMM1 cells

Preparation Method

 OMM1 cells were cultured in RPMI-1640 medium, supplemented with 10% fetal bovine serum (FBS), and 1% penicillin/streptomycin, at 37°C in an incubator with 5% CO2. Cells were seeded into 96-well plates at a density of 3×104 cells/ml and incubated for 24h at 37°C. Cells were exposed to gradient concentrations of CCCP (0, 0.1, 1, 10, and 100µM) for 72h, cell viability was measured.

Reaction Conditions

 0, 0.1, 1, 10, and 100µM; 72h

Applications

 CCCP treatment reduced cell viability in OMM1 cells in a dose-dependent manner.
Animal experiment [2]:

Animal models

 Male C57BL/6J mice

Preparation Method

Male C57BL/6J mice (8 weeks old; 23-25g) were housed under standard conditions with ad libitum access to food and normal drinking water. After one week of routine feeding to acclimate to the environment, the mice underwent one week of preoperative adaptive exercise training and then underwent intracerebral hemorrhage surgery to induce an ICH model. Mice were fasted for 12 h and deprived of water for 4h prior to anesthesia. Initial anesthesia was induced using 1.5% isoflurane and maintained at 1% throughout the procedure. The mice were positioned in a prone orientation on a stereotaxic frame. A small hole (approximately 1mm in diameter) was drilled at the coordinates of the right striatum (0.8mm anterior to bregma, 2mm to the right of the midline, and 3.5mm deep). A solution containing 0.75U of type IV collagenase and 7U/µl heparin in PBS (0.4µl) was injected at a rate of 0.2µl/min over 5min. The needle was left in place for an additional 10min following the injection before withdrawal. The scalp was then sutured, and the mice were allowed to recover in a warm, ventilated environment. In the Sham group, an equal volume of saline was injected, with all other procedures remaining identical. CCCP and Mdivi-1 were diluted with DMSO and injected intraperitoneally at doses of 1mg/kg and 25mg/kg, respectively. 21 days later, the brain tissues of the mice were collected for analysis.

Dosage form

1mg/kg/day; 21 days; i.p.

Applications

CCCP treatment improved neurological functions after ICH in mice.

References:

[1] Xie M, Gu X, Zhao Z, et al. Artemisinin synergizes with CCCP in autophagic cell death induction via ER stress in uveal melanoma[J]. Iscience, 2025, 28(8).

[2] Xiaoyu L, Dandan L, Tianzhao O, et al. Resolvin D1 combined with exercise rehabilitation alleviates neurological injury in mice with intracranial hemorrhage via the BDNF/TrkB/PI3K/AKT pathway[J]. Scientific Reports, 2024, 14(1): 31447.

Product Documents

Purity:>99.50%Appearance:A solid
COASDSData Sheet

Biological Data

1 / 1

Chemical Properties

CAS No.
555-60-2
Synonyms
Carbonyl cyanide m-chlorophenyl hydrazone, NSC 88124
Chemical Name
(3-chlorophenyl)carbonohydrazonoyl dicyanide
SMILES
ClC1=CC(N/N=C(C#N)/C#N)=CC=C1
Molecular Formula
C9H5ClN4
Molecular Weight
204.62
Solubility
≥ 20.5mg/mL in DMSO
Storage
Store at 2-8°C
Shipping Condition
Evaluation samples are shipped with blue ice. Other available sizes are shipped at room temperature or with blue ice upon request.

CalculatorsMolarity / Dilution / Molecular Weight / Unit Conversion / In Vivo Formulation / Solubility

g/mol

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