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Dual-emission dye fluoresces either green or orange, depending upon mitochondrial membrane potential status
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Highly sensitive detection of mitochondrial membrane potential decline
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Aids in the drug research process in preclinical drug safety assessment (ADME-Tox)
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Suitable for chemical/environmental toxicity screening and characterization
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True mix-and-read homogeneous assay for live cells
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Optimized for fluorescence microscopy and flow cytometry
Cell-based assays for evaluating the functional status of mitochondria are emerging as useful tools for elucidating the role of mitochondrial activity in drug-induced toxicity, the apoptosis cascade and other cellular and biochemical processes. The loss of the mitochondrial membrane potential (MMP) is often associated with early stages of apoptosis. The collapse of MMP 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.
MITO-ID® Membrane Potential Detection Kit measures mitochondrial membrane potential with a cationic dye that fluoresces either green or orange depending upon membrane potential status. In energized cells, the MITO-ID® Membrane Potential reagent exists as a green fluorescent monomer in the cytosol and also accumulates as orange-fluorescent aggregates in the mitochondria. However, in cells with compromised mitochondrial membrane potential, the MITO-ID® Membrane Potential reagent exists primarily as green-fluorescent monomers throughout the cytosol and no longer exhibits orange fluorescence in the mitochondria.
Figure 1: The mitochondria of HeLa cells were stained with MITO-ID® Membrane Potential reagent, and visualized by epifluorescence microscopy. Orange fluorescent aggregates are localized in the mitochondria (Orange channel), while green fluorescent monomers mainly stain the cytosol (FITC channel).
Figure 2: Flow Cytometric Analysis of Control and Treated Cells. Jurkat cells were untreated (left) and were treated with 1 μM CCCP for 15 mins (right). Cells were then stained with Enzo MITO-ID® Membrane Potential Dye, and run on a FACS Calibur instrument.
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Product Specification
| Applications: | Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS
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| Application Notes: | The MITO-ID® Membrane Potential Detection Kit has been optimized for measurement of mitochondria membrane potential (MMP) and cell viability by conventional fluorescence microscopy and flow cytometry. |
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| Quality Control: | - % purity of MITO-ID® MP Detection Reagent by TLC ≥ 95%
- A sample kit from each lot of MITO-ID® Membrane Potential Detection Kit for fluorescence microscopy and flow cytometry is assayed using a microplate-based assay. The Z’-factor from CCCP-treated cells is greater than 0.6.
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| Handling: | Protect from light. Avoid freeze/thaw cycles. |
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| Shipping: | Shipped on Dry Ice |
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| Short Term Storage: | -20°C |
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| Long Term Storage: | -80°C |
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| Contents: | MITO-ID® MP Detection Reagent
Necrosis Detection Reagent
CCCP Control
10X Assay Buffer 1
50X Assay Buffer 2 |
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| Technical Info/Product Notes: | The MITO-ID® Membrane potential detection kit is a member of the CELLESTIAL® product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications. CELLESTIAL® reagents and kits are optimal for use in demanding cell analysis applications involving confocal microscopy, flow cytometry, microplate readers and HCS/HTS, where consistency and reproducibility are required.
Toxicology Application Note:
Use of 3D Cultured Human iPSC-Derived Hepatocytes for Long-Term Hepatotoxicity Studies |
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Product Literature References
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Oxidative Stress Induced by Pt(IV) Pro-drugs Based on the Cisplatin Scaffold and Indole Carboxylic Acids in Axial Position: D. Tolan, et al.; Sci. Rep.
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Diallyl trisulfide induces apoptosis in Jurkat cells by the modification of cysteine residues in thioredoxin: K. Watanabe, et al.; Biosci. Biotechnol. Biochem.
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Application(s): Flow cytometry measurements on Jurkat cells,
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Expanding the clinical phenotypes of MT-ATP6 mutations: E. Lopez-Gallardo, et al.; Hum. Mol. Genet.
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Proteome variations in pancreatic stellate cells upon stimulation with pro-inflammatory factors: A.J. Marzog, et al.; J. Biol. Chem.
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Application(s): Apoptotis assay in pancreatic stellate cells,
Abstract;
Full Text
Utilization of fluorescent probes for the quantification and identification of subcellular proteomes and biological processes regulated by lipid peroxidation products: T.D. Cummins, et al.; Free Radic. Biol. Med.
59, 56 (2013),
Application(s): Cell Culture,
Abstract;
Full Text
CPTH6, a thiazole derivative, induces histone hypoacetylation and apoptosis in human leukemia cells: D. Trisciuoglio, et al.; Clin. Cancer Res.
18, 475 (2012),
Application(s): Detection of membrane potential in AML and solid tumor cell lines by flow cytometry,
Abstract;
Full Text
Docosahexaenoic acid-induced unfolded protein response, cell cycle arrest, and apoptosis in vascular smooth muscle cells are triggered by Ca²⁺-dependent induction of oxidative stress: S. Crnkovic, et al.; Free Radic. Biol. Med.
52, 1786 (2012),
Application(s): Detection of membrane potential in human pulmonary artery smooth muscle cells by flow cytometry,
Abstract;
Full Text
Ectopic ATP synthase blockade suppresses lung adenocarcinoma growth by activating the unfolded protein response: H.Y. Chang, et al.; Cancer Res.
72, 4696 (2012),
Application(s): Mitochondrial membrane potential in lung carcinoma cells by fluorescence microscopy,
Abstract;
Full Text
Phenytoin reduces 5-aminolevulinic acid-induced protoporphyrin IX accumulation in malignant glioma cells: M. Hefti, et al.; J. Neurooncol.
108, 443 (2012),
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General Literature References
High throughput fluorescence assays for the measurement of mitochondrial activity in intact human neuroblastoma cells: A.J. Woollacott & P.B. Simpson; J. Biomol. Screen
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Abstract;
A New Method for the Cytofluorometric Analysis of Mitochondrial Membrane Potential Using the J-Aggregate Forming Lipophilic Cation 5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine Iodide (JC-1): A. Cossarizza, et al.; Biochem. Biophys. Res. Commun.
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Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate forming lipophilic cation JC-1: S.T. Smiley, et al.; PNAS
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