For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    jc-10-mitochondrial-membrane-potential-assay-kit-flow-cytometry-ab112133.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Cell Biology Apoptosis Mitochondrial
Share by email

JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (5)References (4)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Flow Cytometry - JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133)

    Key features and details

    • Assay type: Direct
    • Detection method: Fluorescent
    • Platform: Flow cytometer
    • Assay time: 20 min
    • Sample type: Adherent cells, Suspension cells

    You may also be interested in

    Biochemical
    Product image
    FCCP, mitochondrial oxidative phosphorylation uncoupler (ab120081)
    Assay
    Product image
    Glycerol-3-Phosphate Dehydrogenase (G3PDH) Assay Kit (Colorimetric) (ab174095)
    ELISA
    Product image
    Human Frataxin ELISA Kit (ab176112)

    View more associated products

    Overview

    • Product name

      JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry)
      See all Mitochondrial Membrane Potential kits
    • Detection method

      Fluorescent
    • Sample type

      Adherent cells, Suspension cells
    • Assay type

      Direct
    • Assay time

      0h 20m
    • Product overview

      JC-10 Mitochondrial Membrane Potential Assay Kit ab112133 is designed for use with flow cytometry, and it provides the most robust assay method for monitoring changes in mitochondrial membrane potential.


      The 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, changes to a monomeric form and stains cells with 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. Compared to JC-1, JC-10 has much better water solubility.


      JC-10 selectively enters mitochondria, and reversibly changes its color from green to orange-red as membrane potentials increase. This property is due to the reversible formation of JC-10 aggregates upon membrane polarization which cause a shifts in emitted light from 520 nm (the emission of JC-10 monomeric form) to 570 nm (the emission of JC-10-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). Both colors can be detected using the filters commonly mounted in all flow cytometers. Besides its use in flow cytometry, it can also be used in fluorescence imaging and fluorescence microplate platform.


      JC-10 assay protocol summary:
      - add JC-10 staining solution to experimentally treated cells
      - incubate cells for 15-60 min
      - analyze wth flow cytometer

    • Notes

      If you would like to use JC-10 on a microplate reader, we recommend JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134).

      Related assays

      Review the cell health assay guide to learn about kits to perform a cell viability assay, cytotoxicity assay and cell proliferation assay. 

      Review the metabolism assay guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

    • Platform

      Flow cytometer

    Properties

    • Storage instructions

      Store at -20°C. Please refer to protocols.
    • Components 100 tests
      200X JC-10 in DMSO 1 x 250µl
      Assay Buffer A 1 x 50ml
    • Research areas

      • Cell Biology
      • Apoptosis
      • Mitochondrial
      • Kits/ Lysates/ Other
      • Kits
      • Apoptosis Kits
      • Other Apoptosis Kits
      • Kits/ Lysates/ Other
      • Kits
      • Cell Metabolism Kits
      • Cell Viability and Senescence Kits
      • Kits/ Lysates/ Other
      • Kits
      • Cell Damage Kits
      • Cell viability, plasma membrane damage
      • Metabolism
      • Pathways and Processes
      • Metabolism processes
      • Apoptosis
      • Kits/ Lysates/ Other
      • Kits
      • Cell Damage Kits
      • Cell Damage
      • Kits/ Lysates/ Other
      • Kits
      • Apoptosis Kits
      • Transmembrane potential
      • Cancer
      • Cell Death
      • Apoptosis
      • Mitochondrial
      • Cancer
      • Cell Death
      • Apoptosis
      • Metabolism
      • Metabolism
      • Pathways and Processes
      • Mitochondrial Metabolism
      • Membrane potential
    • Relevance

      Mitochondrial Membrane Potential is an important parameter of mitochondrial function used as an indicator of cell death. The collapse of the mitochondrial Membrane potential coincides with the opening of the mitochondrial permeability transition pores, leading to the release of cytochrome c into the cytosol, which in turn triggers other downstream events in the apoptotic cascade.
    • Alternative names

      • mitochondrial membrane potential

    Associated products

    • Assay kits

      • JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134)
      • JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850)
      • TMRE-Mitochondrial Membrane Potential Assay Kit (ab113852)
    • Related Products

      • JC-1, Mitochondrial membrane potential dye (ab141387)

    Images

    • Flow Cytometry - JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133)
      Flow Cytometry - JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133)

      JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133) was used to measure the effect of FCCP induced mitochondria membrane potential change in Jurkat cells by Flow Cytometry. Jurkat cells were dye loaded with JC-10 dye-loading solution along with DMSO (Top) or 5 µM FCCP (Low) for 10 minutes. The fluorescent intensities for both J-aggregates and monomeric forms of JC-10 were measured with a flow cytometer using FL1 and FL2 channels. Uncompensated data (left column) were compared with compensated data (right column).

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (4)

    Publishing research using ab112133? Please let us know so that we can cite the reference in this datasheet.

    ab112133 has been referenced in 4 publications.

    • Bellanti F  et al. Inhibition of nuclear factor (erythroid-derived 2)-like 2 promotes hepatic progenitor cell activation and differentiation. NPJ Regen Med 6:28 (2021). PubMed: 34039998
    • Condelli V  et al. Targeting TRAP1 as a downstream effector of BRAF cytoprotective pathway: a novel strategy for human BRAF-driven colorectal carcinoma. Oncotarget 6:22298-309 (2015). PubMed: 26084290
    • Khurana S  et al. Antiapoptotic actions of methyl gallate on neonatal rat cardiac myocytes exposed to H2O2. Oxid Med Cell Longev 2014:657512 (2014). Flow Cyt ; Rat . PubMed: 24672637
    • Liu Z  et al. Potent Half-Sandwich Iridium(III) Anticancer Complexes Containing C(?)N-Chelated and Pyridine Ligands. Organometallics 33:5324-5333 (2014). PubMed: 25328266

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-5 of 5 Abreviews or Q&A

    Question

    To whom it may concern,
    All our FACS samples are fix using BD cytofix/ctyoperm, I just want to make sure that the JC-10 mitochondrial membrane potential Assay kit (Ab112133) is compatible with fixation/permeabilization.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 03 2014

    Answer





    This kit is for live cells only.



    Read More

    Heather Allen

    Abcam Scientific Support

    Answered on Sep 03 2014

    Question

    I was having troubles with the JC-10 kit I had ordered from your company when using it for flow cytometry.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 18 2013

    Answer

    Based on the data, it looks like the way your customer treated the cells probably is the reason for the problem. It most likely when detaching the cells (trypsinized?), the mitochondria membrane potential changes. If you have to use flow, might try EDTA (which at least make the membrane intake, but I am not 100% sure if it will also cause the same problem) to lift the cells from plate. The best way for the adherent cell mitochondria membrane potential measurement will be the plate format (you do need have a bottom read mode fluorescent plate reader though). In this way, one can keep the cell intake before the experiment.
    Also, due to the great solubility of the JC-10, the sensitivity of the JC-10 on mitochondria membrane potential is very high, especially for the the cells which are very sensitive to environment. It is probably also one of the causes of these "unexpected" results. The following links are some examples that people use JC-10 in cardiomyocytes. Attached please also find a JC-10 paper published last Nov.

    http://www.cellulardynamics.com/products/lit/CDI_appnote_Cardiomyocytes_mitochondrial.pdf


    http://www.moleculardevices.com/Documents/general-documents/scientific-posters/imaging-posters/2011Poster_SBS%20Cardiotoxicity%20Imaging.pdf

    Here are some tips for flow cytometry:
    1. Lift cells with EDTA only (no trypsin), hopefully this procedure will not change the mitochondria membrane potential.
    2. treat FCCP for 15 min in growth medium first.
    3. remove the FCCP.
    4. Resuspend cells in growth medium,
    5. add equal volume of JC-10 dye-loading solution.
    6. incubate for 15 min
    7. analyze with flow.

    Read More

    Abcam Scientific Support

    Answered on Jan 18 2013

    Question

    I chose 10uM FCCP because in your protocol given it says to use between 2-10UM of CCCP or FCCP. I decided to try 10uM because I have previously done JC-1 experiments with an Invitrogen kit and they recommended a concentration of 50uM so I just chose the highest concentration you suggested...

    Read More

    Abcam community

    Verified customer

    Asked on Dec 17 2012

    Answer



    The concentration of CCCP or FCCP required as a positive control will vary based on cell line. Perhaps it would be useful to try 50uM FCCP as this has worked for you in the past.

    Read More

    Abcam Scientific Support

    Answered on Dec 17 2012

    Question

    I apologize for the late email but I have attached data from my JC-10 experiment. My main question is why CCCP is giving a greater FL2/FL1 ratio than my stained control? I thought the reverse was true?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 09 2012

    Answer

    Thank you for providing that extra information.

    I have talked to the lab again, to see if they have any other thoughts on this and they think it is probably due to using the different "gating method", attached please find a copy of the method that was used for gating.

    Yes, you can dye load for 30 min first, and then treat with CCCP for 15 min, we have used FCCP for our positive control.

    If changing the gating does not prove to be successful for you, then I would be happy to either replace or refund the kit for you.

    Please let me know if there is anything I can help you with.

    Read More

    Abcam Scientific Support

    Answered on Nov 09 2012

    Question

    I apologize for the late email but I have attached data from my JC-10 experiment. My main question is why CCCP is giving a greater FL2/FL1 ratio than my stained control? I thought the reverse was true?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 05 2012

    Answer

    Thank you for providing that information.

    I have talked to our supplier about your concerns and they have told me that your results are most likely due to the method of preparing the cells. Physically scraped the cells may damage the cell membrane already. You can tell there are two populations from Control, one with much lower F1 intensity which I believe is more healthy cells, and after CCCP treatment, that population is gone. I recommend you use EDTA (without trypsin) to dissociated the cells from plate.

    Please let me know if there is anything else I can help you with.

    Read More

    Abcam Scientific Support

    Answered on Nov 06 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Get resources and offers direct to your inbox Sign up
    A-Z by research area
    • Cancer
    • Cardiovascular
    • Cell biology
    • Developmental biology
    • Epigenetics & Nuclear signaling
    • Immunology
    • Metabolism
    • Microbiology
    • Neuroscience
    • Signal transduction
    • Stem cells
    A-Z by product type
    • Primary antibodies
    • Secondary antibodies
    • Biochemicals
    • Isotype controls
    • Flow cytometry multi-color selector
    • Kits
    • Loading controls
    • Lysates
    • Peptides
    • Proteins
    • Slides
    • Tags and cell markers
    • Tools & Reagents
    Help & support
    • Support
    • Make an Inquiry
    • Protocols & troubleshooting
    • Placing an order
    • RabMAb products
    • Biochemical product FAQs
    • Training
    • Browse by Target
    Company
    • Corporate site
    • Investor relations
    • Company news
    • Careers
    • About us
    • Blog
    Events
    • Tradeshows
    • Conferences
    International websites
    • abcam.cn
    • abcam.co.jp

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    • Modern slavery statement
    © 1998-2023 Abcam plc. All rights reserved.