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  1. Link

    cccp-mitochondrial-oxidative-phosphorylation-uncoupler-ab141229.pdf

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Biochemicals Chemical Type Biochemicals
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CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)

  • Datasheet
  • SDS
  • COA
Submit a review Submit a question References (11)

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Chemical Structure - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
  • Immunocytochemistry/ Immunofluorescence - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
  • Western blot - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
  • Immunoprecipitation - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
  • Functional Studies - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)

Key features and details

  • Potent mitochondrial oxidative phosphorylation uncoupler
  • CAS Number: 555-60-2
  • Purity: > 99%
  • Soluble in DMSO to 100 mM and in ethanol to 100 mM
  • Form / State: Solid
  • Source: Synthetic

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Overview

  • Product name

    CCCP, Mitochondrial oxidative phosphorylation uncoupler
  • Description

    Potent mitochondrial oxidative phosphorylation uncoupler
  • Biological description

    Potent mitochondrial oxidative phosphorylation uncoupler. Renders mitochondrial inner membrane permeable to protons. Induces apoptosis in vitro.

  • Purity

    > 99%
  • CAS Number

    555-60-2
  • Chemical structure

    Chemical Structure

Properties

  • Chemical name

    2-[2-(3-Chlorophenyl)hydrazinylyidene]propanedinitrile
  • Molecular weight

    204.62
  • Molecular formula

    C9H5ClN4
  • PubChem identifier

    2603
  • Storage instructions

    Store at -20°C. Store under desiccating conditions. The product can be stored for up to 12 months.
  • Solubility overview

    Soluble in DMSO to 100 mM and in ethanol to 100 mM
  • Handling

    Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one month. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.

    Toxic, refer to SDS for further information.

    Need more advice on solubility, usage and handling? Please visit our frequently asked questions (FAQ) page for more details.

  • SMILES

    Clc1cc(N\N=C(/C#N)C#N)ccc1
  • Source

    Synthetic

  • Research areas

    • Biochemicals
    • Chemical Type
    • Biochemicals
    • Biochemicals
    • Pharmacology
    • Signaling
    • Apoptosis & cell cycle
    • Other
    • Biochemicals
    • Research Area
    • Cancer
    • Apoptosis & cell cycle
    • Other

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab141229 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies
Use at an assay dependent concentration.
Notes
Functional Studies
Use at an assay dependent concentration.

Images

  • Chemical Structure - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
    Chemical Structure - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
    2D chemical structure image of ab141229, CCCP, Mitochondrial oxidative phosphorylation uncoupler
  • Immunocytochemistry/ Immunofluorescence - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
    Immunocytochemistry/ Immunofluorescence - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)

    Immunofluorescent analysis of 4 % paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma)(+/- treatment with 10μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) cells labeling PINK1 with ab216144 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells treated with 10μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    The negative controls are as follows:
    -ve control: PBS, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Western blot - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
    Western blot - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
    All lanes : Anti-PINK1 antibody [EPR20730] (ab216144) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa cells (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Observed band size: 62 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking and dilution buffer: 5% NFDM/TBST

    PINK1 can be induced by CCCP treatment (PMID: 24184327).

  • Immunoprecipitation - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
    Immunoprecipitation - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)

    PINK1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP. ab141229) for 24 hours) whole cell lysate with ab216144 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216144 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: HeLa (CCCP-treated, ab141229) lysate 10 μg (Input). 
    Lane 2: ab216144 IP in HeLa (CCCP-treated, ab141229) lysate. 
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216144 in HeLa (CCCP-treated, ab141229) whole cell lysate.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Functional Studies - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)
    Functional Studies - CCCP, Mitochondrial oxidative phosphorylation uncoupler (ab141229)

    MCF7 cells were incubated at 37°C for 2 hours with vehicle control (0 μM) and different concentrations of CCCP (ab 141229). Increased expression of AKT1 (phospho S473) (ab81283) in MCF7 cells correlates with an increase in CCCP concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10 μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab81283 at 2 μg/ml and ab8227 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download
  • COA

References (11)

Publishing research using ab141229? Please let us know so that we can cite the reference in this datasheet.

ab141229 has been referenced in 11 publications.

  • Voigt AL  et al. Unique metabolic phenotype and its transition during maturation of juvenile male germ cells. FASEB J 35:e21513 (2021). PubMed: 33811704
  • Hu LL  et al. Functional role and molecular mechanisms underlying prohibitin 2 in platelet mitophagy and activation. Mol Med Rep 23:N/A (2021). PubMed: 33760146
  • Orabi KY  et al. Psiadin and plectranthone selectively inhibit colorectal carcinoma cells proliferation via modulating cyclins signaling and apoptotic pathways. PLoS One 16:e0252820 (2021). PubMed: 34086816
  • Kara E  et al. An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of a-synuclein. Cell Rep 35:109189 (2021). PubMed: 34107263
  • Kim C  et al. SUMOylation of mitofusins: A potential mechanism for perinuclear mitochondrial congression in cells treated with mitochondrial stressors. Biochim Biophys Acta Mol Basis Dis 1867:166104 (2021). PubMed: 33617988
View all Publications for this product

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