Cell Meter™ NIR Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
Quotation | Request |
International | See distributors |
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
Cell Meter™ NIR Mitochondrion Membrane Potential Assay Kit *Optimized for Microplate Reader* |
Related products
Overview | ![]() ![]() |
Our Cell Meter™ assay kits are a set of tools for monitoring cell viability. There are a variety of parameters that can be used for monitoring cell viability. This particular kit is designed to monitor cell apoptosis through measuring the loss of the mitochondrial membrane potential. 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™ NIR Membrane Potential Detection Kit provides all the essential components with an optimized assay method for the detection of apoptosis in cells with the loss of mitochondrial membrane potential. This fluorometric assay is based on the detection of the mitochondrial membrane potential in cells by our proprietary cationic MitoLite NIR™ dye. In normal cells, MitoLite NIR™ accumulates primarily in mitochondria, however, in apoptotic cells, MitoLite NIR™ staining intensity decreases. Cells stained with MitoLite NIR™ can be visualized by flow cytometry with red excitation and far red emission (FL4 channel). The kit can be paired with other reagents, such as blue-excited propidium iodide and Cell Meter™ Phosphatidylserine Apoptosis Assay Kit (#22803) for multi-parametric study of cell vitality and apoptosis. The kit is optimized for screening of apoptosis activators and inhibitors by flow cytometry.
Platform
Flow cytometer
Excitation | 640 nm laser |
Emission | 660/20 nm filter |
Instrument specification(s) | APC 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 5 µL of 200X MitoLite™ NIR 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 FL4 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 MitoLite™ NIR. To get the best result, titration of the CCCP or FCCP may be required for each individual cell line. - Add 5 µL of 200X MitoLite™ NIR (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 MitoLite™ NIR 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 FL4 channel (Ex/Em = 635/660 nm). Gate on the cells of interest, excluding debris.
Images

Figure 1. The decrease in fluorescence intensity of MitoLite™ NIR with the addition of FCCP in Jurkat cells. Jurkat cells were loaded with MitoLite™ NIR alone (blue line) or in the presence of 50 µM FCCP (red line) for 10 minutes. The fluorescence intensity of MitoLite™ NIR was measured with a FACSCalibur (Becton Dickinson) flow cytometer using FL4 channel.
References
View all 91 references: Citation Explorer
Safranine O as a fluorescent probe for mitochondrial membrane potential studied on the single particle level and in suspension
Authors: Perevoshchikova IV, Sorochkina AI, Zorov DB, Antonenko YN.
Journal: Biochemistry (Mosc) (2009): 663
Authors: Perevoshchikova IV, Sorochkina AI, Zorov DB, Antonenko YN.
Journal: Biochemistry (Mosc) (2009): 663
Determination of high mitochondrial membrane potential in spermatozoa loaded with the mitochondrial probe 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) by using fluorescence-activated flow cytometry
Authors: Guthrie HD, Welch GR.
Journal: Methods Mol Biol (2008): 89
Authors: Guthrie HD, Welch GR.
Journal: Methods Mol Biol (2008): 89
The mitochondrial membrane potential and Ca2+ oscillations in smooth muscle
Authors: Chalmers S, McCarron JG.
Journal: J Cell Sci (2008): 75
Authors: Chalmers S, McCarron JG.
Journal: J Cell Sci (2008): 75
Computer-assisted live cell analysis of mitochondrial membrane potential, morphology and calcium handling
Authors: Koopman WJ, Distelmaier F, Esseling JJ, Smeitink JA, Willems PH.
Journal: Methods (2008): 304
Authors: Koopman WJ, Distelmaier F, Esseling JJ, Smeitink JA, Willems PH.
Journal: Methods (2008): 304
How DASPMI reveals mitochondrial membrane potential: fluorescence decay kinetics and steady-state anisotropy in living cells
Authors: Ramadass R, Bereiter-Hahn J.
Journal: Biophys J (2008): 4068
Authors: Ramadass R, Bereiter-Hahn J.
Journal: Biophys J (2008): 4068
Life cell quantification of mitochondrial membrane potential at the single organelle level
Authors: Distelmaier F, Koopman WJ, Testa ER, de Jong AS, Swarts HG, Mayatepek E, Smeitink JA, Willems PH.
Journal: Cytometry A (2008): 129
Authors: Distelmaier F, Koopman WJ, Testa ER, de Jong AS, Swarts HG, Mayatepek E, Smeitink JA, Willems PH.
Journal: Cytometry A (2008): 129
Effects of eprosartan on mitochondrial membrane potential and H2O2 levels in leucocytes in hypertension
Authors: Labios M, Martinez M, Gabriel F, Guiral V, Ruiz-Aja S, Beltran B, Munoz A.
Journal: J Hum Hypertens (2008): 493
Authors: Labios M, Martinez M, Gabriel F, Guiral V, Ruiz-Aja S, Beltran B, Munoz A.
Journal: J Hum Hypertens (2008): 493
Mitochondrial membrane potential in axons increases with local nerve growth factor or semaphorin signaling
Authors: Verburg J, Hollenbeck PJ.
Journal: J Neurosci (2008): 8306
Authors: Verburg J, Hollenbeck PJ.
Journal: J Neurosci (2008): 8306
Evaluation of sperm mitochondrial membrane potential by JC-1 fluorescent staining and flow cytometry
Authors: Xia XY, Wu YM, Hou BS, Yang B, Pan LJ, Shi YC, Jin BF, Shao Y, Cui YX, Huang YF.
Journal: Zhonghua Nan Ke Xue (2008): 135
Authors: Xia XY, Wu YM, Hou BS, Yang B, Pan LJ, Shi YC, Jin BF, Shao Y, Cui YX, Huang YF.
Journal: Zhonghua Nan Ke Xue (2008): 135
Cyclosporin A-induced oxidative stress is not the consequence of an increase in mitochondrial membrane potential
Authors: van der Toorn M, Kauffman HF, van der Deen M, Slebos DJ, Koeter GH, Gans RO, Bakker SJ.
Journal: Febs J (2007): 3003
Authors: van der Toorn M, Kauffman HF, van der Deen M, Slebos DJ, Koeter GH, Gans RO, Bakker SJ.
Journal: Febs J (2007): 3003
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