2. Neuronal Signaling Autophagy Others Immunology/Inflammation Metabolic Enzyme/Protease Anti-infection Apoptosis
  3. Oxidative Phosphorylation PINK1/Parkin STING IFNAR Mitochondrial Metabolism Bacterial Apoptosis
  4. CCCP

CCCP  (Synonyms: Carbonyl cyanide 3-chlorophenylhydrazone; Carbonyl Cyanide m-Chlorophenylhydrazone)

Cat. No.: HY-100941 Purity: 98.75%
COA Handling Instructions

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CCCP is an oxidative phosphorylation (OXPHOS) uncoupler. CCCP induces activation of PINK1 leading to Parkin Ser65 phosphorylation.

For research use only. We do not sell to patients.

CCCP Chemical Structure

CCCP Chemical Structure

CAS No. : 555-60-2

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10 mM * 1 mL in DMSO
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Customer Review

Based on 68 publication(s) in Google Scholar

CCCP Related Antibodies

Top Publications Citing Use of Products

68 Publications Citing Use of MCE CCCP

  • Biological Activity

  • Protocol

  • Purity & Documentation

  • References

  • Customer Review

Description

CCCP is an oxidative phosphorylation (OXPHOS) uncoupler. CCCP induces activation of PINK1 leading to Parkin Ser65 phosphorylation[1].

IC50 & Target

STING[1]
IFN-β[1]
In Vitro

CCCP inhibits IFN-β production induced by various types of the STING pathway activators. CCCP suppresses the phosphorylation of STING, TBK1, and IRF3 via disrupting the association of STING and TBK1. CCCP inhibits activation of STING and its downstream signaling molecules, TBK1 and IRF3, but not STING translocation to the perinuclear region. CCCP impairs the interaction between STING and TBK1 and concomitantly triggers mitochondria fission. Importantly, the knockout of the crucial mitochondria fission regulator Drp1 restored the STING activity, indicating that CCCP down-modulates the STING pathway through DRP1-mediated mitochondria fragmentation. The protonophore CCCP that disrupts membrane potential suppresses the DMXAA-triggered STING signaling pathway. CCCP drastically suppresses the production of IFN-β in DMXAA-treated RAW264.7 cells and MEFs[1].
As low as 1 μM CCCP is enough to induce mitocytosis. In cells treated with 10 μM CCCP, which is the dose used for inducing mitophagy, mitocytosis is barely induced. Mechanistically, mitocytosis requires positioning of damaged mitochondria at the cell periphery, which occurs because damaged mitochondria avoid binding to inward motor proteins[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

CCCP Related Antibodies
In Vivo

The same dosage of 3 mg/kg.bw each of CCCP and PPEF is used. In both the cases 1 log reduction is observed in the bacterial load. However, when 3 mg/kg.bw of PPEF is used in combination with 3 mg/kg.bw of CCCP, 6 log10 reduction is observed in the bacterial count. The developed model validates the enhanced antibacterial activity of combination therapy[2].99mTc-MIBI signals in the hearts of SD rats administered CCCP (4 mg/kg intraperitoneally) or vehicle is also measured. 99mTc-MIBI signals decrease in rat hearts administered CCCP, and the ATP content, as measured by 31P magnetic resonance spectroscopy, decreased simultaneously. To investigate whether CCCP decreased the 99mTc-MIBI signals in rats, we analyzed the radioisotope activity of excised heart tissue from rats administered CCCP. At 180 min after 99mTc-MIBI injection, the 99mTc-MIBI signals from the hearts in the CCCP group are significantly lower than those in the vehicle group[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Molecular Weight

204.62

Formula

C9H5ClN4
CAS No.
Appearance

Solid

Color

Yellow to brown

SMILES

N#C/C(C#N)=N/NC1=CC=CC(Cl)=C1
Shipping

Room temperature in continental US; may vary elsewhere.
Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 2 years
-20°C 1 year
Solvent & Solubility
In Vitro: 

DMSO : 50 mg/mL (244.36 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : < 0.1 mg/mL (insoluble)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 4.8871 mL 24.4355 mL 48.8711 mL
5 mM 0.9774 mL 4.8871 mL 9.7742 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (12.22 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
% 
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 99.83%

COA (245 KB) HNMR (163 KB) LCMS (386 KB)

Handling Instructions (2659 KB)
References
Cell Assay
[1]

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].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
Animal Administration
[2][3]

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).

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 4.8871 mL 24.4355 mL 48.8711 mL 122.1777 mL
5 mM 0.9774 mL 4.8871 mL 9.7742 mL 24.4355 mL
10 mM 0.4887 mL 2.4436 mL 4.8871 mL 12.2178 mL
15 mM 0.3258 mL 1.6290 mL 3.2581 mL 8.1452 mL
20 mM 0.2444 mL 1.2218 mL 2.4436 mL 6.1089 mL
25 mM 0.1955 mL 0.9774 mL 1.9548 mL 4.8871 mL
30 mM 0.1629 mL 0.8145 mL 1.6290 mL 4.0726 mL
40 mM 0.1222 mL 0.6109 mL 1.2218 mL 3.0544 mL
50 mM 0.0977 mL 0.4887 mL 0.9774 mL 2.4436 mL
60 mM 0.0815 mL 0.4073 mL 0.8145 mL 2.0363 mL
80 mM 0.0611 mL 0.3054 mL 0.6109 mL 1.5272 mL
100 mM 0.0489 mL 0.2444 mL 0.4887 mL 1.2218 mL
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CCCP Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

CCCP555-60-2Carbonyl cyanide 3-chlorophenylhydrazone Carbonyl Cyanide m-ChlorophenylhydrazoneOxidative PhosphorylationPINK1/ParkinSTINGIFNARMitochondrial MetabolismBacterialApoptosisStimulator of Interferon GenesTMEM173MITAERISMPYSInterferon-α/β receptorInterferon-alpha/beta receptor Inhibitorinhibitorinhibit

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