Skip to main content

JC1- Mitochondrial Membrane Potential Assay Kit ab113850 contains tetraethylbenzimidazolylcarbocyanine iodide (JC-1), a cationic dye that accumulates in energized mitochondria.


Images

Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850), expandable thumbnail
  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850), expandable thumbnail
  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850), expandable thumbnail
  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850), expandable thumbnail
  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850), expandable thumbnail

Publications

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay time

1h

Associated Products

Select an associated product type

4 products for Alternative Product

What's included?

100 Test
Components
50mM FCCP (in DMSO)
1 x 10 µL
DMSO (cell culture tested)
1 x 1 mL
Dilution Buffer (10X, sterile)
1 x 10 mL
JC-1 (lyophilized)
1 x 500 µg

Recommended products

JC1- Mitochondrial Membrane Potential Assay Kit ab113850 contains tetraethylbenzimidazolylcarbocyanine iodide (JC-1), a cationic dye that accumulates in energized mitochondria.

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay time

1h

Assay Platform

Microplate reader

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

Multi

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

JC1- Mitochondrial Membrane Potential Assay Kit ab113850 contains tetraethylbenzimidazolylcarbocyanine iodide (JC-1), a cationic dye that accumulates in energized mitochondria.

At low concentrations (due to low mitochondrial membrane potential), JC-1 is predominantly a monomer that yields green fluorescence with emission of 530±15 nm.

At high concentrations (due to high mitochondrial membrane potential), the dye aggregates yielding a red to orange colored emission (590±17.5 nm).

Therefore a decrease in the aggregate fluorescent count is indicative of depolarization whereas an increase is indicative of hyperpolarization.

The JC-1 staining protocol is very simple:
- wash cells in dilution buffer or PBS
- add JC solution
- incubate for 30 min at 37°C for suspension cells, or 10 min for adherent cells
- wash cells with dilution buffer
- treat cells as desired for experimental plan
- analyze on a fluorescent microplate reader

The aggregate dye can be excited at 535 nm, the monomer and aggregate together at 475 nm.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The mitochondrial membrane potential also known as ΔΨm is the electrical potential difference across the inner mitochondrial membrane. This potential results from the electrochemical gradient produced by the proton pumps during electron transport chain activity. The mechanical function of the mitochondrial membrane potential is important to ATP production through oxidative phosphorylation. Mitochondrial membranes are widely expressed in almost all eukaryotic cells and are an essential component of cellular metabolism. The inner membrane is structured to facilitate its function housing integral proteins that are key to maintaining the potential.

Biological function summary

The mitochondrial membrane potential drives ATP synthesis by powering ATP synthase an enzyme complex embedded in the mitochondrial membrane. This potential also plays a vital role in other processes such as calcium homeostasis and regulation of mitochondrial biogenesis. The mitochondrial membrane itself forms part of the larger mitochondrial respiratory chain complex coordinating with components like complex I (NADH: ubiquinone oxidoreductase) and complex II (succinate dehydrogenase) to maintain cell energy needs and respond to metabolic demands.

Pathways

The mitochondrial membrane potential is integral to cellular energy metabolism pathways such as the Krebs cycle and oxidative phosphorylation. Mitochondrial membrane potential modulation can affect signaling proteins like cytochrome c which is instrumental in apoptosis. Apoptotic signaling pathways involving proteins such as Bax and Bcl-2 influence the mitochondrial membrane potential and regulate cell survival or death in response to cellular stress or damage.

Associated diseases and disorders

Changes in the mitochondrial membrane potential relate significantly to conditions like neurodegenerative diseases and cancer. In neurodegenerative diseases such as Parkinson’s and Alzheimer's dysregulation of mitochondrial membrane potential can lead to impaired energy production and increased oxidative stress. Cancer cells often exhibit altered mitochondrial membrane potential affecting processes like apoptosis and enabling survival in adverse conditions. These alterations in potential impact proteins such as p53 which play critical roles in cancer progression and neurodegenerative disease pathology.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

6 product images

  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850), expandable thumbnail

    Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850)

    JC-1 assay result in HL60 cells treated with FCCP.

    JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850). HL60 cells were seeded and labeled according to section 11.1 of the protocol. Cells were then treated for 4 hours with 100 μM FCCP or vehicle/diluent control (DMSO). Mean and standard deviation is plotted for 3 replicates from each condition.

  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850), expandable thumbnail

    Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850)

    JC-1 assay result in HepG2 cells treated with CCCP.

    JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850). HepG2 cells were seeded and labeled according to section 11.2 of the protocol. Cells were then treated for 4 hours with a titration series of CCCP (carbonyl cyanide 3-chlorophenylhydrazone) and both monomer and aggregate forms were read on a Perkin Elmer-Wallac 1420 Victor 2 Multilabel plate reader. Mean and standard deviation of aggregate/monomer ratios is plotted for 12 replicates for each concentration. IC50 of CCCP in HepG2 cells was calculated at 8.7 μM.

  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850), expandable thumbnail
    Image courtesy of Son M S et al. Sci Rep. 2017; 7: 2075. doi: 10.1038/s41598-017-02129-w. Reproduced under the Creative Commons License http://creativecommons.org/licenses/by/4.0/.

    Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850)

    Using JC1 dye to examine mitochondrial membrane potential depolarization in SH-SY5Y cells.

    Son MS et al. (2017) used JC1 Mitochondrial Membrane Potential Assay Kit ab113850 to stain:
    - untreated SH-SY5Y cells (control),
    - SH-SY5Y cells treated with 1mM MPP+ (MPP+) and,
    - SH-SY5Y cells pretreated with BDS-II followed by 1mM MPP+ treatment (MPP+ +BDSII).

    Normal mitochondrial membrane potential is shown in red with JC-1 dimers and depolarized membrane potential is shown in green in JC-1 monomers.

  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850), expandable thumbnail

    Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850)

    JC-1 assay result in HL60 cells treated with Troglitazone.

    JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850). HL60 cells were seeded and labeled according to section 11.1 of the protocol. Cells were then treated for 4 hours with a titration series of the thiazolidinedione Troglitazone and both monomer and aggregate forms were read on a Perkin Elmer-Wallac 1420 Victor 2 Multilabel plate reader. Mean and standard deviation of aggregate/monomer ratios is plotted for 3 replicates for each concentration. IC50 of Troglitazone in HL60 cells was calculated at 1.2 μM.

  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850), expandable thumbnail
    This image is courtesy of a customer review submitted by Dimitra Kalamida

    Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850)

    JC1 Mitochondrial Membrane Potential Assay Kit using green fluorescence imaging.

    The JC1- Mitochondrial membrane potential assay kit has been tested using HepG2 cells, control cells and FCCP-treated cells (100uM for 4h) have been used as a positive control. The company's instructions were followed for JC1 mitochondrial membrane potential assay. Imaging was performed on a customized Andor Revolution Spinning Disk Confocal System built around a stand (IX81 Olympus) with a 60x lens and a digital camera (Andor Ixon+885) (CIBIT Facility, MBG-DUTH). Image acquisition was performed in Andor IQ 2 software. Optical sections were recorded every 0.3 μm. All confocal microscopy images presented in this work are 2D maximum intensity projections of z-stack images (ImageJ 1.47v National Institute of Health,USA).
    Personal feedback: A green laser with the appropriate emission filter (530nm) has been used to detect the monomer of the JC1 dye, following FCCP treatment the mitochondrial membrane potential of the cells was eliminated, as demonstrated by the increase of the monomer emission.

  • Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850), expandable thumbnail
    Image courtesy of Koukourakis M I et al. Sci Rep. 2016; 6: 30986doi: 10.1038/srep30986. Reproduced under the Creative Commons License http://creativecommons.org/licenses/by/4.0/

    Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850)

    JC1 used to assay in vitro metabolic effect on hepatocyte and hepatoma cells.

    Koukourakis MI et al. (2016) used the MMP assay kit to stain with JC-1 in NCTC hepatocytes exposed to amifostine (100 μg/ml) over a time course of 20 minutes in vitro. Confocal microscopy used to assess mitochondrial membrane potential in the cells.

    Serial images confirmed a rapid drop of both green and red fluorescence, one minute after exposure, an effect that was restored to normal at 20 minutes, after a small period of a rebound increase.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com