Description
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.
Product details:
How the assay works
The TMRE Mitochondrial Membrane Potential Assay Kit uses a fluorescent dye called tetramethylrhodamine ethyl ester (TMRE).
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 assay 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.
How other researchers are using TMRE assay kit ab113852
TMRE-Mitochondrial Membrane Potential Assay Kit has been used in a variety of sample type including:
Human pulmonary arterial endothelial cells (PAECs)
Mice spermatozoa
Human adipose-derived mesenchymal stem cells (AD-MSCs)
References:
1-Piper B et al. 2024
2-Park S al. 2024
3-Abu-El-Rub E et al. 2023
Related and recommended products
See other alternative kits to quantify Mitochondrial Membrane Potential:
- JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate)
ab112134
- JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry)
ab112133
- JC-1 - Mitochondrial Membrane Potential Assay Kit
ab113850
Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.
