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MitoSciences (ms960)

TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)

Q&A (13)Specific References (1)

Overview

  • Product nameTMRE—Mitochondrial Membrane Potential Assay KitSee all Mitochondrial Transmembrane Potential kits ...
  • Product overview

    ab113852 is suitable for interrogating mitochondrial membrane potential in live cells for analysis by flow cytometry, microplate spectrophotometry and fluorescent microscopy. Each assay kit contains sufficient materials for at least 200 measurements.

    ab113852 uses TMRE (tetramethylrhodamine, ethyl ester) 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.

    FCCP (carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone) 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.

  • Tested applicationsFunctional Studies more details

Properties

  • RelevanceMitochondrial Transmembrane Potential is an important parameter of mitochondrial function used as an indicator of cell death. The collapse of the mitochondrial transmembrane 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 potentialMS960

    Applications

    Our Abpromise guarantee covers the use of ab113852 in the following tested applications.

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

    Application Notes
    Functional Studies FuncS

    TMRE—Mitochondrial Membrane Potential Assay Kit images

    • 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.
    • 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.
    • Functional Studies - TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)
      Functional Studies - TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)
      Microplate analysis of cells showing mean fluorescent intensity +/- standard deviation from quadruplicate measurements of 400nM ab113852 (TMRE) stained Jurkat cells in a 96-well microplate +/- treatment with FCCP.

    Protocols

    References for TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852)

    This product has been referenced in:
    • Jandova J  et al. Cyclophilin 40 alters UVA-induced apoptosis and mitochondrial ROS generation in keratinocytes. Exp Cell Res 319:750-60 (2013). Read more (PubMed: 23220213) »

    See 1 Publication for this product

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

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    Submit a Question for ab113852

    Displaying 1 - 10 of 13 results for Q&A

    10
    Votes: Highest First

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


    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...

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    I am trying to use this kit as follows:
    Samples: mouse embryonic fibroblasts plated at two different densities
    20 minute incubation with 100-500 nM TMRE diluted in media
    Read in 0.2% BSA at ex 525, em 580-640 nm
    I am getting RFU ˜ 10^2...

    Read More

    Thank you for contacting us. My colleague from our MitoSciences lab has provided the following recommendations.

    The lack of signal may be due to working outside the peak excitation (549nm) and peak emission (575nm). Getting as close as possible ...

    Read More

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

    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) i...

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    I have blood smears with red blood cells infected by intracellular parasites that grow in RPMI medium. These slides are already done and they are in storage for a few months. I am looking for mitochondria markers to evaluate apoptosis by fluorescence micr...

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    Thank you for contacting us.

    I have received the following information from the lab:

    TMRE can only be used with live cells—an active membrane potential is required for staining. This will be true of all membrane potential sensit...

    Read More

    Hello,

    I'm wondering about what solutionI can use to dilute TMRE? Would PBS containing 0.2% BSA be fine?

    I used DMSO and the assay is not working, probably because high amount od DMSO was too toxic for the cells.

    Thank you for your inquiry.

    Culture media is known to cause background using kit ab113852. For suspension cells, we recommend pelleting the cells, removing the culture media, resuspending in the same volume of 0.2% BSA in PBS, pelleting again, a...

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    I have an inquiry regarding your TMRE—Mitochondrial Membrane Potential Assay Kit (ab113852).

    I'm planning to do the microscopy assay on a very sensitve cell type that do not adhere to a glass slide.

    I'm wondeing if Icanseed and...

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    Thank you for contacting us.

    Culture media is known to cause background using kit ab113852. For suspension cells, we recommend pelleting the cells, removing the culture media, resuspending in the same volume of 0.2% BSA in PBS, pelleting again, ...

    Read More

    Dear abcam technical support team: This customer has highly interested in ab113849 (Luminescent ATP Detection Assay Kit), ab113850 (JC1 - Mitochondrial Membrane Potential Assay Kit) and ab113852 (TMRE—Mitochondrial Membrane Potential Assay Kit), thi...

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    Thank you for contacting us. We have not tested these kits on yeast samples; however there are papers in the literature showing the use of TMRE in yeast as well as measurement of ATP using luciferin. See below title of paper and methods details. Attention...

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    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?

    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 distinc...

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    Thank you very much for your reply, I forwarded the mail to the customer. She now explains her project and wants me to ask you if you have any ideas, or products which she might be able to use.   Here is her mail:   Thanks so much for finding out f...

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    Thank you for your reply. I have been in touch with our colleagues at Mitosciences who have kindly provided the following information: To answer your question directly, The antibody coating the plate supplied with Kit MTOX1 ab109903 does not cross...

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     I am interested in your mitochondria isolation kit for cultured cells. In the product description you mention that isolated mitochondria can be analyzed by activity assays. Do you sell also specific assay kits, or can me tell which activity assay, idealy...

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    Thank you very much for your question about our mitochondria isolation kit ab110170 . I have consulted our specialist from Mitosciences in order to be able to provide you with the best possible answer - please find them here below: Our mitochondrial isola...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"