製品の概要

  • 製品名
    TMRE-Mitochondrial Membrane Potential Assay Kit
    Mitochondrial Transmembrane Potential キット 製品一覧
  • 検出方法
    Fluorescent
  • アッセイタイプ
    Cell-based (qualitative)
  • 製品の概要

    TMRE-Mitochondrial Membrane Potential Assay Kit (ab113852) is suitable for quantifying changes in mitochondrial membrane potential in live cells 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.


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

  • アプリケーション
    適用あり: Functional Studiesmore details
  • 試験プラットフォーム
    Reagents

法規制情報

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
Functional Studies Use at an assay dependent concentration.

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  • 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.

  • 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.
  • 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 histogram of Jurkat cells stained with ab113852 (100nM TMRE) with (blue) or without (red) treatment with 100µM FCCP.

プロトコール

参考文献

This product has been referenced in:
  • Chen HH  et al. Immune dysregulation in patients with PTEN hamartoma tumor syndrome: Analysis of FOXP3 regulatory T cells. J Allergy Clin Immunol 139:607-620.e15 (2017). Functional Studies ; Human . Read more (PubMed: 27477328) »
  • Zhang K & Jiang D RhoA inhibits the hypoxia-induced apoptosis and mitochondrial dysfunction in chondrocytes via positively regulating the CREB phosphorylation. Biosci Rep 37:N/A (2017). Read more (PubMed: 28254846) »

See all 22 Publications for this product

レビューと Q&A

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...

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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 dist...

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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.







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

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 kn...

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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.

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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 possib...

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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 sen...

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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...

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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 agai...

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1-10 of 16 Abreviews or Q&A

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

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