JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134)


  • Product name
    JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate)
    See all Mitochondrial Membrane Potential kits
  • Detection method
  • Sample type
    Adherent cells, Suspension cells
  • Assay type
  • Product overview

    JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134) enables researchers to run JC-10 assay in the format of microplate reader, and it provides the most robust assay method for monitoring mitochondria membrane potential changes. This assay is based on the detection of the mitochondrial membrane potential changes in cells by the cationic, lipophilic JC-10 dye. In normal cells, JC-10 concentrates in the mitochondrial matrix where it forms red fluorescent aggregates. However, in apoptotic and necrotic cells, JC-10 diffuses out of mitochondria. It changes to monomeric form and stains cells in green fluorescence. 

    Although JC-1 is widely used in many labs, its poor water solubility causes great inconvenience. Even at 1 µM concentration, JC-1 tends to precipitate in aqueous buffer. JC-10 is developed to be a superior alternative to JC-1 when high dye concentration is desired. Compared to JC-1, JC-10 has much better water solubility. JC-10 is capable of selectively entering mitochondria, and reversibly changes its color from green to orange as membrane potentials increase. This property is due to the reversible formation of JC-10 aggregates upon membrane polarization that causes shifts in emitted light from 520 nm (i.e. emission of JC-10 monomeric form) to 570 nm (i.e. emission of J-aggregate form). When excited at 490 nm, the color of JC-10 changes reversibly from green to greenish orange as the mitochondrial membrane becomes more polarized. 

    In normal cells, JC-10 concentrates in the mitochondrial matrix where it forms red fluorescent aggregates. However, in apoptotic and necrotic cells, JC-10 exists in monomeric form and stains cells green. The green emission can be analyzed in fluorescence channel 1 (FL1) and greenish orange emission in channel 2 (FL2). Besides its use in fluorescence microplate platform, it can also be used in fluorescence imaging and flow cytometry.

     Visit our FAQs page for tips and troubleshooting.

    Review our cell health assays guide to learn more about our other cell viability, cytotoxicity and cell proliferation assay kits.

  • Notes

    A microplate reader with bottom-reading mode is essential to perform this assay.

    If you would like to use JC-10 on a flow cytometer, we recommend JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133).

  • Platform



  • CGN were cultured on 96-well white-walled clear-bottom plates in phenol-red free Neurobasal until 7 DIV. Thirty minutes before the end of the treatment, 50 μl of JC-10 dye-loading solution was added to each well and incubated for 30 minutes before measuring fluorescence intensities (Ex/Em  = 485/515 nm and Ex/Em  = 540/590 nm). The change of mitochondrial membrane potential was measured as the ratio between aggregate (Em  = 515 nm) and monomeric forms (Em  = 590 nm) of JC-10. Increasing ratios indicate mitochondrial membrane depolarization.

  • JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134). Camptothecin-induced mitochondria membrane potential changes with JC-10 and JC-1 in Jurkat cells. JC-1 (blue bar) and JC-10 (red bar) dye loading solutions were added to Jurket cells untreated (0 µM) or treated with camptothecin (10 µM for 4 hours) and incubated for 30 minutes. The fluorescent intensities for both J-aggregates and monomeric forms of JC-1 and JC-10 were measured at Ex/Em = 490/525 nm and 490/590 nm with a microplate reader.



This product has been referenced in:
  • Wang D & Liu P Ingenol-3-Angelate Suppresses Growth of Melanoma Cells and Skin Tumor Development by Downregulation of NF-?B-Cox2 Signaling. Med Sci Monit 24:486-502 (2018). Human . Read more (PubMed: 29368698) »
  • Li S  et al. Mitochondrial Dysfunctions Contribute to Hypertrophic Cardiomyopathy in Patient iPSC-Derived Cardiomyocytes with MT-RNR2 Mutation. Stem Cell Reports 10:808-821 (2018). Human . Read more (PubMed: 29456182) »

See all 11 Publications for this product

Customer reviews and Q&As

Thank you for your enquiry.

JC-1, a lipophiliccationic dye, is widely used to detect mitochondrial de-polarization. However it has poor water solubility which means it precipitates in aqueous buffer even at 1 μM concentration. Compared to J...

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You will not remove the treatment media prior to step C3.
The final volume in the well is 200 uL (100 uL cells + 50 uL JC-10 + 50 uL assay buffer B).

Assay Buffer B is used to quench the reaction and reduce the overall background. It is not necessary to perform the assay successfully however, it will definitely improve the results.

Phenol red should be OK; in our experience there has not been any interference.

The concentration of 100X JC-10 in ab112134 is 3 mM. The solubility is ˜ 5mg/mL in DMSO.

I can confirm that the numbers seems to be alright to me.

The product is used for mitochondria membrane potential measurement.

The dye needs to get into the live cells to exhibit either green (in cytosol) or red (in mitoch...

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Thank you for contacting us about ab112134. Here is some further information: - The blank contains no cells and is growth media only. This gives the background and should be subtracted from all readings. - The control sample should have cells wi...

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Yes, you can do it, although the background might increase a bit.

Thank you for your message.

I understand the confusion and apologize for the inconvenience.

We have checked this and the documents have been amended with the correct information: We recommend to incubate at either room temperature...

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Q1. Are clear bottomed wells required for plates used in this assay?
A1. Yes

Q2. Serum (1%) will be present in medium when JC10 is added. Will JC10 be effected by serum?
A2. No
Q3. Is there an optimal height setting at which t...

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1-10 of 18 Abreviews or Q&A


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