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| Product Name | Mitochondrial Membrane Potential Fluorescence Assay Kit (TMRE) |
| Catalog No. | CCAT-HMM-0053 |
| Description | A rapid fluorescence-based assay using tetramethylrhodamine ethyl ester (TMRE), a cell-permeable cationic fluorescent dye that accumulates in active mitochondria in proportion to the mitochondrial inner membrane potential. TMRE is chemically stable and exhibits minimal photobleaching compared to JC-1, with a reversible potential-dependent uptake mechanism. |
| Intended Use | Real-time monitoring of mitochondrial membrane potential in live cells for mitochondrial toxicity screening, metabolic flux studies, and assessment of early apoptotic events preceding caspase activation. |
| Principle / Technology | TMRE (Ex/Em: 549/574 nm), a lipophilic cation with delocalized positive charge, distributes across the mitochondrial inner membrane according to the Nernst equation; in cells with high ΔΨm (approximately -180 mV), TMRE accumulates in the mitochondrial matrix at concentrations 400–500× the external medium; mitochondrial depolarization leads to TMRE redistribution into the cytoplasm, decreasing the overall fluorescence intensity in the quenching mode or altering signal in non-quenching mode. |
| Detection Method | Fluorescence microscopy with TRITC/Rhodamine filter set (Ex/Em: 549/574 nm), flow cytometry (488 nm or 561 nm laser), or fluorescence plate reader |
| Sample Type | Live adherent and suspension mammalian cells; isolated mitochondria; primary neurons, cardiomyocytes, and hepatocytes for mitochondrial toxicity testing |
| Performance Range / Specifications | Staining concentration: 20–100 nM for non-quenching mode, 1–2 μM for quenching mode; incubation 15–30 minutes at 37°C; reversible potential response within seconds of uncoupler addition; chemical stability allows repeated imaging over 2–3 hours |
| Sensitivity / LOD | Detection of ΔΨm changes induced by 0.5 μM FCCP within 1–2 minutes of treatment; resolves potential differences of approximately 5–10 mV |
| Specificity | TMRE uptake is specific to membranes with high negative-inside potential difference; mitochondrial inner membrane under normal conditions (-150 to -180 mV) drives selective accumulation; minimal binding to plasma membrane or endoplasmic reticulum at standard concentrations; FCCP/CCCP fully dissipates TMRE gradient as positive control |
| Reaction Conditions / Protocol | Prepare TMRE working solution at desired concentration in cell culture medium or imaging buffer; replace medium with TMRE solution; incubate 15–30 minutes at 37°C under 5% CO2; wash once if using non-quenching mode, or retain dye if using quenching mode; image with TRITC filter or analyze by flow cytometry; include FCCP-treated cells as depolarization positive control |
| Components / Formulation | TMRE (tetramethylrhodamine, ethyl ester, perchlorate) supplied as 1 mg lyophilized or 10 mM DMSO stock; DMSO for reconstitution; FCCP positive control solution (optional); detailed protocol with guidance on quenching versus non-quenching mode selection |
| Storage Conditions | Lyophilized TMRE: -20°C protected from light and desiccated, stable 24 months; DMSO stock (10 mM): -20°C protected from light, desiccated, stable 6 months; working solution: fresh use only |
| Shelf Life | 24 months from date of manufacture for lyophilized form |
| Package Specifications | 25 mg, 100 mg lyophilized; 1 × 10 mM DMSO solution |
| Product Form | Lyophilized solid or concentrated DMSO stock solution |
| Quality Control | Each lot tested for mitochondrial staining specificity using HeLa cells ± 10 μM FCCP; mean fluorescence intensity ratio (untreated/FCCP) ≥3; purity ≥95% by HPLC |
| Key Features | Low photobleaching rate compared to JC-1 and DiOC6(3); compatible with real-time kinetic imaging for dynamic potential changes; quenching mode at high concentration provides self-referencing signal; works with standard TRITC/Rhodamine filter cubes on all fluorescence microscopes |
For research use only, not for clinical use.
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