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.
Images
Publications
- The Journal of clinical investigation 134:2024RAB7 deficiency impairs pulmonary artery endothelial function and promotes pulmonary hypertension.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBryce Piper,Srimathi Bogamuwa,Tanvir Hossain,Daniela Farkas,Lorena Rosas,Adam C Green,Geoffrey Newcomb,Nuo Sun,Jose A Ovando-Ricardez,Jeffrey C Horowitz,Aneel R Bhagwani,Hu Yang,Tatiana V Kudryashova,Mauricio Rojas,Ana L Mora,Pearlly Yan,Rama K Mallampalli,Elena A Goncharova,David M Eckmann,Laszlo FarkasPubMed 38015641
- Reproductive biology and endocrinology : RB&E 22:152024Inhibition of mitochondrial cyclophilin D, a downstream target of glycogen synthase kinase 3α, improves sperm motility.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSeung Hyun Park,Myung Chan GyePubMed 38254112
Key facts
- Detection method
Fluorescent
- Sample types
Suspension cells, Adherent cells
- Assay type
Cell-based (qualitative)
- Assay time
35m
Associated Products
Select an associated product type
5 products for Alternative Product
What's included?
Recommended products
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.
Key facts
- Detection method
Fluorescent
- Sample types
Suspension cells, Adherent cells
- Assay type
Cell-based (qualitative)
- Assay time
35m
- Assay Platform
Microplate reader, Fluor. microscope, Flow cyt.
Storage
- Shipped at conditions
Blue Ice
- Appropriate long-term storage conditions
+4°C
- Storage information
+4°C
Notes
Store kit at 4°C 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 Tetramethylrhodamine, ethyl ester 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.
TMRE is only suitable for use with live (not fixed) cells. **Related assays** Review the to learn about kits to perform a , and . Review the 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
Supplementary info
- Activity summary
The mitochondrial membrane potential also known as ΔΨm is the electrical potential difference across the inner mitochondrial membrane. This potential results from the electrochemical gradient produced by the proton pumps during electron transport chain activity. The mechanical function of the mitochondrial membrane potential is important to ATP production through oxidative phosphorylation. Mitochondrial membranes are widely expressed in almost all eukaryotic cells and are an essential component of cellular metabolism. The inner membrane is structured to facilitate its function housing integral proteins that are key to maintaining the potential.
- Biological function summary
The mitochondrial membrane potential drives ATP synthesis by powering ATP synthase an enzyme complex embedded in the mitochondrial membrane. This potential also plays a vital role in other processes such as calcium homeostasis and regulation of mitochondrial biogenesis. The mitochondrial membrane itself forms part of the larger mitochondrial respiratory chain complex coordinating with components like complex I (NADH: ubiquinone oxidoreductase) and complex II (succinate dehydrogenase) to maintain cell energy needs and respond to metabolic demands.
- Pathways
The mitochondrial membrane potential is integral to cellular energy metabolism pathways such as the Krebs cycle and oxidative phosphorylation. Mitochondrial membrane potential modulation can affect signaling proteins like cytochrome c which is instrumental in apoptosis. Apoptotic signaling pathways involving proteins such as Bax and Bcl-2 influence the mitochondrial membrane potential and regulate cell survival or death in response to cellular stress or damage.
- Associated diseases and disorders
Changes in the mitochondrial membrane potential relate significantly to conditions like neurodegenerative diseases and cancer. In neurodegenerative diseases such as Parkinson’s and Alzheimer's dysregulation of mitochondrial membrane potential can lead to impaired energy production and increased oxidative stress. Cancer cells often exhibit altered mitochondrial membrane potential affecting processes like apoptosis and enabling survival in adverse conditions. These alterations in potential impact proteins such as p53 which play critical roles in cancer progression and neurodegenerative disease pathology.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
6 product images
Popova 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/Functional Studies - TMRE-Mitochondrial Membrane Potential Assay Kit (ab113852)
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)
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.
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.
Image courtesy of Maiti A K et al. PLoS One. 2018; 13(9): e0204567. doi: 10.1371/journal.pone.0204567Functional Studies - TMRE-Mitochondrial Membrane Potential Assay Kit (ab113852)
Mitochondrial membrane potential assayed using TMRE in the study of VIP.
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.
Functional Studies - TMRE-Mitochondrial Membrane Potential Assay Kit (ab113852)
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.
Image 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/Functional Studies - TMRE-Mitochondrial Membrane Potential Assay Kit (ab113852)
TMRE assay to assess mitochondrial function after C. rodentium infection.
Maiti et al (2015) used TMRE assay kit ab113852 to assess mitochondrial membrane potential in murine distal colon after C. rodentium infection.
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com