Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Orange Fluorescence Optimized for Flow Cytometry*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Alternative formats
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See also: Flow Cytometry Reagents
This particular kit is designed to detect cell apoptosis by measuring the loss of the mitochondrial membrane potential(MMP). The collapse of mitochondrial membrane potential 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. Our Cell Meter™ Orange Mitochondrial Membrane Potential Assay Kit provides all the essential components with an optimized assay method. This fluorimetric assay uses our proprietary cationic MitoLite™ Orange for the detection of apoptosis in cells with the loss of mitochondrial membrane potential. In normal cells, the red fluorescence intensity is increased when MitoLite™ Orange is accumulated in the mitochondria. However, in apoptotic cells, the fluorescence intensity of MitoLite™ Orange is decreased following the collapse of MMP. Cells stained with MitoLite™ Orange can be visualized with a flow cytometer at 488 nm excitation with red emission (FL2 channel). The kit can be used together with other reagents, such as Cell Meter™ Phosphatidylserine Apoptosis Assay Kit (22835) for multi-parametric study of cell vitality and apoptosis. The kit is optimized for screening apoptosis activators and inhibitors with a flow cytometer.
Platform
Flow cytometer
Excitation | 488 nm or 532 nm laser |
Emission | 575/26 nm filter |
Instrument specification(s) | PE channel |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells with test compounds at the density of 5 × 105 to 1 × 106 cells/mL
- Add 2 µL of 500X MitoTell™ Orange into 1 mL of cell solution
- Incubate the cells in a 37 °C, 5% CO2 incubator for 15 - 30 minutes
- Pellet the cells, and resuspend the cells in 1 mL of growth medium
- Analyze cells using flow cytometer with FL2 channel
Important notes
Thaw all the kit components at room temperature before starting the experiment.
SAMPLE EXPERIMENTAL PROTOCOL
- For each sample, prepare cells in 1 mL of warm medium or buffer of your choice at the density of 5×105 to 1×106 cells/mL. Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density for apoptosis induction.
- Treat cells with test compounds for a desired period of time to induce apoptosis, and set up parallel control experiments.
For Negative Control: Treat cells with vehicle only.
For Positive Control: Treat cells with FCCP or CCCP at 5-50 µM in a 37 oC, 5% CO2 incubator for 15 to 30 minutes. Note: CCCP or FCCP can be added simultaneously with MitoTell™ Orange. To get the best result, titration of the CCCP or FCCP may be required for each individual cell line. - Add 2 µL of 500X MitoTell™ Orange (Component A) into the treated cells.
- Incubate the cells in a 37 °C, 5% CO2 incubator for 15 to 30 minutes. Note: For adherent cells, gently lift the cells with 0.5 mM EDTA to keep the cells intact and wash the cells once with serum-containing media prior to the incubation with MitoTell™ Orange dye-loading solution.
- Centrifuge the cells at 1000 rpm for 4 minutes, and then re-suspend cells in 1 mL of Assay Buffer (Component B) or buffer of your choice.
- Monitor the fluorescence intensity using a flow cytometer wih FL2 channel (Ex/Em = 540/590 nm). Gate on the cells of interest, excluding debris.
Product Family
Name | Excitation (nm) | Emission (nm) |
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Red Fluorescence Optimized for Flow Cytometry* | 613 | 631 |
Images

Figure 1. The decrease in fluorescence intensity of MitoTell™ Orange with the addition of FCCP in Jurkat cells. Jurkat cells were loaded with MitoTell™ Orange alone (Blue) or in the presence of 30 µM FCCP (Red) for 15 minutes. The fluorescence intensity of MitoTell™ Orange was measured with a FACSCalibur (Becton Dickinson, San Jose, CA) flow cytometer using FL2 channel.
Citations
View all 2 citations: Citation Explorer
Evaluation of the oncolytic property of recombinant Newcastle disease virus strain R2B in 4T1 and B16-F10 cells in-vitro
Authors: Ramamurthy, Narayan and Pathak, Dinesh C and D'Silva, Ajai Lawrence and Batheja, Rahul and Mariappan, Asok Kumar and Vakharia, Vikram N and Chellappa, Madhan Mohan and Dey, Sohini
Journal: Research in Veterinary Science (2021): 159--165
Authors: Ramamurthy, Narayan and Pathak, Dinesh C and D'Silva, Ajai Lawrence and Batheja, Rahul and Mariappan, Asok Kumar and Vakharia, Vikram N and Chellappa, Madhan Mohan and Dey, Sohini
Journal: Research in Veterinary Science (2021): 159--165
Mitochondrial Dysfunction Contributes to Aging-Related Atrial Fibrillation
Authors: Liu, Chuanbin and Bai, Jing and Dan, Qing and Yang, Xue and Lin, Kun and Fu, Zihao and Lu, Xu and Xie, Xiaoye and Liu, Jianwei and Fan, Li and others,
Journal: Oxidative Medicine and Cellular Longevity (2021)
Authors: Liu, Chuanbin and Bai, Jing and Dan, Qing and Yang, Xue and Lin, Kun and Fu, Zihao and Lu, Xu and Xie, Xiaoye and Liu, Jianwei and Fan, Li and others,
Journal: Oxidative Medicine and Cellular Longevity (2021)
References
View all 91 references: Citation Explorer
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Journal: J Neurosci (2008): 8306
Evaluation of sperm mitochondrial membrane potential by JC-1 fluorescent staining and flow cytometry
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Journal: Zhonghua Nan Ke Xue (2008): 135
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