GLPBio

FCCP

Catalog No.: GC14328Purity:>98.00%Synonyms: Trifluoromethoxy carbonylcyanide phenylhydrazone
FCCP is a highly lipophilic mitochondrial uncoupler.

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FCCP
Cas No.: 370-86-5
10mM (in 1mL DMSO)In Stock
$55.00
1
10mgIn Stock
$50.00
1
50mgIn Stock
$190.00
1

Citations

4 publicationsPublications from researchers using this product

Description

FCCP is a highly lipophilic mitochondrial uncoupler. FCCP acts as a protonophore across the inner mitochondrial membrane to transport protons, dissipate mitochondrial membrane potential and pH gradient, uncouple electron transport from ATP synthesis, increase oxygen consumption while inhibiting ATP production, and trigger energy stress by reducing mitochondrial ATP synthesis, affecting AMPK and other energy sensing nodes as well as HIF signaling. FCCP can be used for research on mitochondrial function, cellular energy metabolism, hypoxia signaling and tumor metabolism[1-4].

In vitro, co-treatment of HuH6, Hep3B, HepG2 and HLE cells with 1μM FCCP and Gemcitabine for 16-48 hours inhibited Gemcitabine-induced mitochondrial superoxide generation and induced resistance to Gemcitabine-inhibited cell growth[5]. Treatment of niclosamide-pretreated HEK293T cells with 10μM FCCP for 1 hour reduced maximal cellular respiration rate[6]. Treatment of 4T1 or B16 cells with 0.5μM FCCP for 30 minutes induced peak uncoupled oxygen consumption rate[7].

In vivo, twice-daily intraperitoneal injection of 1mg/kg FCCP for 1 day in C57BL/6 mice and PINK1-deficient (PINK1-/-) mice caused Parkin translocation to cardiac mitochondria, accompanied by increased ubiquitination of mitochondrial proteins and increased association of autophagy protein LC3II with mitochondria. Retrograde aortic perfusion of wild-type and PINK1-/- mouse hearts with 100nM FCCP for 5-15 minutes caused rapid Parkin accumulation in the cardiac mitochondrial fraction without causing endogenous PINK1 accumulation at mitochondria[8].

References:

[1] Galkina KV, Finkelberg JM, Markova OV, et al. Protonophore FCCP provides fitness advantage to PDR-deficient yeast cells. J Bioenerg Biomembr. 2020 Oct;52(5):383-395.

[2] Park KS, Jo I, Pak K, 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.

[3] 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.

[4] Brennan JP, Southworth R, Medina RA, et al. Mitochondrial uncoupling, with low concentration FCCP, induces ROS-dependent cardioprotection independent of KATP channel activation. Cardiovasc Res. 2006 Nov 1;72(2):313-21.

[5] Yu G, Liu J, Xu K, et al. Uncoupling protein 2 mediates resistance to gemcitabine-induced apoptosis in hepatocellular carcinoma cell lines. Biosci Rep. 2015;35(4):e00231.

[6] Ng MY, Song ZJ, Tan CH, et al. Structural investigations on the mitochondrial uncouplers niclosamide and FCCP. FEBS Open Bio. 2024;14(6):1057-1071.

[7] Jiang X, Fan Z, Zhang Z, et al. Tumor metabolome remolded by low dose mitochondrial uncoupler elicites robust CD8+ T cell response. Cell Death Discov. 2025;11:291.

[8] Kubli DA, Cortez MQ, Moyzis AG, et al. PINK1 Is Dispensable for Mitochondrial Recruitment of Parkin and Activation of Mitophagy in Cardiac Myocytes. PLoS ONE. 2015 Jun 25;10(6):e0130707.

Protocol

Cell experiment [1]:

Cell lines

HuH6,Hep3B, HepG2, HLE cells (human hepatocellular carcinoma cell line)

Preparation Method

HuH6, Hep3B, HepG2 and HLE cells were maintained in Dulbecco's Modified Eagle Medium supplemented with 10% FBS at 37°C, 5% CO₂. Cells were seeded in 96-well plates and treated with 1μM Gemcitabine alone or in combination with FCCP (4-trifluoro-methoxy-phenyl-hydrazone) for 16h (mitochondrial superoxide assay) or 48h (cell growth inhibition assay). After treatment, mitochondrial superoxide was measured by MitoSox Red (Catalog No.GC68230) and cell growth was measured by MTT (Catalog No.GC14731) assay.

Reaction Conditions

1μM; 16h or 48h

Applications

FCCP co-treatment with Gemcitabine induced resistance to Gemcitabine-induced cell growth inhibition in HuH6, Hep3B, HepG2 and HLE cells. FCCP co-treatment with Gemcitabine prevented Gemcitabine-induced mitochondrial superoxide generation in HuH6, Hep3B, HepG2 and HLE cells.
Animal experiment [2]:

Animal models

Wild-type C57BL/6 mice, PINK1-deficient (PINK1-/-) mice

Preparation Method

Mice were intraperitoneally administered 1mg/kg FCCP in saline twice daily. The final dose was given 1 hour prior to tissue collection. Hearts were excised, retrograde perfused via aorta with Krebs-Henseleit buffer for 5min equilibration then with 100nM FCCP in perfusion buffer for 5min or 15min. Ventricular tissue was used for mitochondrial isolation, Western blot of Parkin, PINK1, ubiquitin, LC3II, Tom20, complex markers, and GAPDH.

Dosage form

1mg/kg; i.p.; twice daily for one day

100nM; cardiac perfusion for 5min or 15min

Applications

FCCP administration increased Parkin levels in the mitochondrial fraction of wild-type and PINK1-/- hearts. FCCP perfusion did not cause endogenous PINK1 accumulation at mitochondria in wild-type hearts at 5min or 15min. FCCP in vivo increased Parkin translocation to mitochondria in wild-type and PINK1-/- hearts, increased ubiquitination of mitochondrial proteins, and increased association of LC3II with mitochondria in wild-type and PINK1-/- hearts.

References:

[1] Yu G, Liu J, Xu K, et al. Uncoupling protein 2 mediates resistance to gemcitabine-induced apoptosis in hepatocellular carcinoma cell lines. Biosci Rep. 2015;35(4):e00231.

[2] Kubli DA, Cortez MQ, Moyzis AG, et al. PINK1 Is Dispensable for Mitochondrial Recruitment of Parkin and Activation of Mitophagy in Cardiac Myocytes. PLoS ONE. 2015 Jun 25;10(6):e0130707.

Product Documents

Purity:>98.00%Appearance:A solid

Chemical Properties

CAS No.
370-86-5
Synonyms
Trifluoromethoxy carbonylcyanide phenylhydrazone
Chemical Name
(4-(trifluoromethoxy)phenyl)carbonohydrazonoyl dicyanide
SMILES
FC(F)(OC1=CC=C(C=C1)N/N=C(C#N)/C#N)F
Molecular Formula
C10H5F3N4O
Molecular Weight
254.17
Solubility
≥ 56.6 mg/mL in DMSO with ultrasonic, ≥ 25 mg/mL in EtOH with ultrasonic
Storage
Store at -20°C
General tips
Select a suitable solvent according to the product solubility data. Once prepared, aliquot the stock solution to avoid repeated freeze-thaw cycles.
Use stock solutions within 6 months when stored at -80°C or within 1 month when stored at -20°C.
To improve solubility, warm the tube to 37°C and sonicate it for an appropriate period.
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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