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Cell Biology Apoptosis Mitochondrial
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TMRE-Mitochondrial Membrane Potential Assay Kit (ab113852)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (4)Q&A (16)References (157)

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Mitochondrial membrane potential was measured using ab113852
  • Immunocytochemistry/ Immunofluorescence - TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)
  • Mitochondrial membrane potential assayed using TMRE in the study of VIP
  • Analysis of TMRE staining using a fluorescent plate reader and a microplate.
  • Flow Cytometry - TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)
  • TMRE assay to assess mitochondrial function after C. rodentium infection

Key features and details

  • Assay type: Cell-based (qualitative)
  • Detection method: Fluorescent
  • Platform: Microplate reader, Fluor. microscope, Flow cyt.
  • Assay time: 35 min
  • Sample type: Adherent cells, Suspension cells

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Overview

  • Product name

    TMRE-Mitochondrial Membrane Potential Assay Kit
    See all Mitochondrial Membrane Potential kits
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Cell-based (qualitative)
  • Assay time

    0h 35m
  • Product overview

    Store kit at 4ºC in the dark immediately upon receipt.


    TMRE-Mitochondrial Membrane Potential Assay Kit ab113852 is used for quantifying changes in mitochondrial membrane potential in live cells by flow cytometry, microplate spectrophotometry and fluorescent microscopy. 


    TMRE (tetramethylrhodamine, ethyl ester) is used to label active mitochondria. TMRE is a cell permeant, positively-charged, red-orange dye that readily accumulates in active mitochondria due to their relative negative charge. Depolarized or inactive mitochondria have decreased membrane potential and fail to sequester TMRE. NB: TMRE is also available as free molecule as ab274305 (Tetramethylrhodamine ethyl ester).


    The TMRE protocol also uses FCCP (carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone), which is a ionophore uncoupler of oxidative phosphorylation. Treating cells with FCCP eliminates mitochondrial membrane potential and TMRE staining. TMRE is suitable for the labeling of mitochondria in live cells and is not compatible with fixation.


    TMRE protocol summary:
    - add FCCP to appropriate control cell samples and incubate for 10 min
    - incubate with TMRE for 15-30 min, pellet (suspension cells) / remove media (adherent cells) and wash with PBS / 0.2% BSA
    - analyze with micro-plate reader at Ex/Em 549/575 nm, flow cytometer using 488nm laser for excitation and at emission 575 nm, or fluorescent microscope.

  • Notes

    TMRE is only suitable for use with live (not fixed) cells.


    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.


    How other researchers have used TMRE assay kit ab113852

    Maiti AK et al. of the University of Gothenburg, Sweden, used TMRE assay kit ab113852 to identify that C. rodentium infection in mice* reduced mitochondrial transmembrane potential. In a subsequent paper, they identified that this effect was reduced by treatment with vasoactive intestinal peptide (VIP). *a model for enteropathogenic E. coli

  • Platform

    Microplate reader, Fluor. microscope, Flow cyt.

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 200 tests
    1mM TMRE (in DMSO) 1 x 40µl
    50mM FCCP (in DMSO) 1 x 10µl
  • 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 (Flow Cytometry) (ab112133)
    • JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134)
    • JC-1 - Mitochondrial Membrane Potential Assay Kit (ab113850)
  • Related Products

    • FCCP, mitochondrial oxidative phosphorylation uncoupler (ab120081)

Images

  • Mitochondrial membrane potential was measured using ab113852
    Mitochondrial membrane potential was measured using ab113852Popova D et al., PloS One, 11(11). Fig 5. doi: 10.1371/journal.pone.0166750 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    P19 neurons (750 cells/mm2) were exposed to MDMA on days 7–9 in serum-free medium for 10 min up to 48 hours. The positive control FCCP (carbonyl cyanide-p-trifluoromethoxyphenylhydrazone), an uncoupler of mitochondrial oxidative phosphorylation, was applied at the concentration of 5 μM for 10 min. The cells were incubated with 500 μM TMRE for 30–45 min at 37°C, 5% CO2, followed by washing once with 100 μl of HBSS containing 0.2% bovine serum albumin. A volume of 200 μL of HBSS containing 0.2% bovine serum albumin was added to each well, and the fluorescence was measured with excitation/emission: 544/590 nm.

  • Immunocytochemistry/ Immunofluorescence - TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)
    Immunocytochemistry/ Immunofluorescence - TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)
    A: HeLa cells (adherent) were cultured on coverslips and stained with ab113852 (200nM TMRE) for 20 minutes in media, washed briefly with PBS and immediately imaged. B: Jurkat cells (suspension) were stained and washed as above and then transferred to a slide and immobilized under a coverslip for imaging.
  • Mitochondrial membrane potential assayed using TMRE in the study of VIP
    Mitochondrial membrane potential assayed using TMRE in the study of VIPImage courtesy of Maiti A K et al. PLoS One. 2018; 13(9): e0204567. doi: 10.1371/journal.pone.0204567

    Maiti AK et al (2018) used TMRE assay kit ab113852 to measure mitochondrial membrane potential in an in vitro mouse intestinal model treated with cytokines in the presence and absence of VIP (vasoactive intestinal peptide). VIP was induced by C. rodentium infection and cytokines.

  • Analysis of TMRE staining using a fluorescent plate reader and a microplate.
    Analysis of TMRE staining using a fluorescent plate reader and a microplate.

    Chart showing mean fluorescent intensity +/- standard deviation from quadruplicate measurements of 400 nM TMRE stained Jurkat cells in a 96-well microplate +/- treatment with FCCP.

  • Flow Cytometry - TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)
    Flow Cytometry - TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)
    Flow cytometry histogram of Jurkat cells stained with ab113852 (100nM TMRE) with (blue) or without (red) treatment with 100µM FCCP.
  • TMRE assay to assess mitochondrial function after C. rodentium infection
    TMRE assay to assess mitochondrial function after C. rodentium infectionImage courtesy of Maiti A K et al. Sci Rep. 2015; 5: 1543. doi: 10.1038/srep15434. Reproduced under the Creative Commons License http://creativecommons.org/licenses/by/4.0/

    Maiti et al (2015) used TMRE assay kit ab113852 to assess mitochondrial membrane potential in murine distal colon after C. rodentium infection.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (157)

Publishing research using ab113852? Please let us know so that we can cite the reference in this datasheet.

ab113852 has been referenced in 157 publications.

  • Li X  et al. Comparison of cytotoxicity effects induced by four different types of nanoparticles in human corneal and conjunctival epithelial cells. Sci Rep 12:155 (2022). PubMed: 34997120
  • Tabebi M  et al. Loss of SDHB Induces a Metabolic Switch in the hPheo1 Cell Line toward Enhanced OXPHOS. Int J Mol Sci 23:N/A (2022). PubMed: 35008989
  • Viheriälä T  et al. Cell maturation influences the ability of hESC-RPE to tolerate cellular stress. Stem Cell Res Ther 13:30 (2022). PubMed: 35073969
  • Afroze N  et al. Fisetin Deters Cell Proliferation, Induces Apoptosis, Alleviates Oxidative Stress and Inflammation in Human Cancer Cells, HeLa. Int J Mol Sci 23:N/A (2022). PubMed: 35163629
  • Perera MR  et al. A Viral Long Non-Coding RNA Protects against Cell Death during Human Cytomegalovirus Infection of CD14+ Monocytes. Viruses 14:N/A (2022). PubMed: 35215840
View all Publications for this product

Customer reviews and Q&As

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

1-10 of 20 Abreviews or Q&A

TMRE using Flow Cytometer

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
The TMRE stain could be detected in PE channel in stained samples in comparison to unstained samples . concentration should be titrated
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 02 2021

TMRE red for mitochondrial membrane potential

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Highly recommended! it just needs to work fast and in the dark.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 14 2020

TMRE red for mitochondrial membrane potential

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Highly recommended! it just needs to work fast and in the dark.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 14 2020

mitochondrial membrane potential in human fibroblast cells

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
To measure mitochondrial activity in human fibroblast cells, we applied TMRE mitochondrial membrane potential assay kit. It showed great correlation with Seahorse analysis which measured OCR (oxygen consumption rate).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 02 2018

Question

What is your recommendation for plates to use with this assay?

Read More

Abcam community

Verified customer

Asked on Jun 14 2012

Answer

Thank you for contacting us. Our lab uses:

BD PureCoat™ Microplates
black/clear, polystyrene
96-well, amine
Cat#354717

The recommendation is for clear bottom, black wall microplate that (a) cells will grow on and (b) is compatible with plate reader.

Please let me know if you have any other questions.

Read More

Abcam Scientific Support

Answered on Jun 14 2012

Question

What are the differences between ab113852 TMRE—Mitochondrial Membrane Potential Assay Kit and ab113850? Which one is more sensitive or more suitable for stem cells?

Read More

Abcam community

Verified customer

Asked on Jan 09 2012

Answer

Thank you for your inquiry. I heard back from the lab regardingthese two distinct dyes that both measure mitochondrial membrane potential. TMRE stains mitochondria only when there is a membrane potential. JC-1 is slightly different in that it has distinct emission spectra depending on whether mitochondria have high or low potential. TMRE can be measured in a spectophotometer, fluorescent microscope or flow cytometry. JC-1 can be measured in a spectrophotometer. Details are in the respective protocol booklets. https://www.abcam.com/ps/products/113/ab113850/documents/ab113850%20Protocol%20(Website)%20v2.pdf https://www.abcam.com/ps/products/113/ab113852/documents/ab113852%20protocol%20final%20v2%20(website(.pdf For you, wewould recommend TMRE with analysis on a flow cytometer. Flow cytometry is both sensitive and amenable to very few cells. I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

Answered on Jan 09 2012

Question

after the TMRE incubation, how long does the user to have to read the samples? Is it immediate, or is there a window after the incubation?

Read More

Abcam community

Verified customer

Asked on Dec 09 2015

Answer

The samples should be read immediately after the 15-30 minute incubation with TMRE. Since the cells are live, as the health of the cell declined the signal from the dye would also diminish.



Read More

Heather Allen

Abcam Scientific Support

Answered on Dec 09 2015

Question



Inquiry: Is there any alternative for a positive control besides FCCP for this assay? Culture condition?

Read More

Abcam community

Verified customer

Asked on Nov 20 2015

Answer





For another positive control, CCCP is suitable (5-50 μM CCCP for 30 to 60 minutes at 37ºC).

Read More

Heather Allen

Abcam Scientific Support

Answered on Nov 20 2015

Question

Can we use this kit to measure mitochondrial membrane potential of yeast (S. cerevisiae or S. pombe)?

Read More

Abcam community

Verified customer

Asked on Oct 02 2015

Answer

It all depends on the sample (embryos vs. explant vs. whole work). We have only worked with this dye using tissue culture cells (monolayer or suspension). If the samples are dissociated cells then TMRE labeling should work. Otherwise, we don’t know how well or deeply the dye can penetrate across cell layers (e.g. into tissues or embryos).


The dye for measuring the potential in the kit is species independent, so it is more a matter of the sample type.

Read More

Kevin Hanson

Abcam Scientific Support

Answered on Oct 02 2015

Question

I have received an e-mail from a customer asking whether AB113852 “is suitable for labelling/staining of mitochondrial fractions extracted from tissues?”

Read More

Abcam community

Verified customer

Asked on Feb 28 2013

Answer


We have no experience in using TMRE to stain isolated mitochondria from any source – it is a tricky proposition as you would need intact/non-damaged mitochondria which retain membrane potential therefore we wouldn't recommend this.

As for imaging tissue stained with TMRE, this is also tricky. Flow cytometry is almost certainly out due to the need for single cell suspension – which will be very difficult for most tissue types. For microscopy I suppose it is possible with the proper microscope set up and the ability to adapt the live tissue to microscope stage constraints (e.g. thin slices.)

However, I would like to stress that this kit is only really for mitochondria labelling in intact cultured cells.

Read More

Abcam Scientific Support

Answered on Feb 28 2013

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