JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133)
- Datasheet
- Specific References (2)
- Protocols
Overview
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Product nameJC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry)
See all Mitochondrial Membrane Potential kits -
Sample typeAdherent cells, Suspension cells
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Assay typeDirect
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Product overview
JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133) enables researchers to run JC-10 assay using flow cytometry, and it provides the most robust assay method for monitoring mitochondria membrane potential changes. This assay is based on the detection of the mitochondrial membrane potential changes in cells by the cationic, lipophilic JC-10 dye. In normal cells, JC-10 concentrates in the mitochondrial matrix where it forms red fluorescent aggregates. However, in apoptotic and necrotic cells, JC-10 diffuses out of mitochondria. It changes to monomeric form and stains cells in green fluorescence.
Although JC-1 is widely used in many labs, its poor water solubility causes great inconvenience. Even at 1 µM concentration, JC-1 tends to precipitate in aqueous buffer. JC-10 is developed to be a superior alternative to JC-1 when high dye concentration is desired. Compared to JC-1, JC-10 has much better water solubility. JC-10 is capable of selectively entering mitochondria, and reversibly changes its color from green to orange as membrane potentials increase. This property is due to the reversible formation of JC-10 aggregates upon membrane polarization that causes shifts in emitted light from 520 nm (i.e., emission of JC-10 monomeric form) to 570 nm (i.e., emission of J-aggregate form). When excited at 490 nm, the color of JC-10 changes reversibly from green to greenish orange as the mitochondrial membrane becomes more polarized.
In normal cells, JC-10 concentrates in the mitochondrial matrix where it forms red fluorescent aggregates. However, in apoptotic and necrotic cells, JC-10 exists in monomeric form and stains cells green. The green emission can be analyzed in fluorescence channel 1 (FL1) and greenish orange emission in channel 2 (FL2). Both colors can be detected using the filters commonly mounted in all flow cytometers. Besides its use in flow cytometry, it can also be used in fluorescence imaging and fluorescence microplate platform.
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Notes
If you would like to use JC-10 on a microplate reader, we recommend JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134).
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Tested applicationsSuitable for: Flow Cytmore details
Properties
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Storage instructionsStore at -20°C. Please refer to protocols.
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Components 100 tests 200X JC-10 in DMSO 1 x 250µl Assay Buffer A 1 x 50ml -
Research areas
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RelevanceMitochondrial Membrane Potential is an important parameter of mitochondrial function used as an indicator of cell death. The collapse of the 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.
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Alternative names
- mitochondrial membrane potential
Associated products
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Assay kits
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Applications
Our Abpromise guarantee covers the use of ab112133 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt | Use at an assay dependent concentration. |
Images
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Flow Cytometry - JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133)JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133) was used to measure the effect of FCCP induced mitochondria membrane potential change in Jurkat cells by Flow Cytometry. Jurkat cells were dye loaded with JC-10 dye-loading solution along with DMSO (Top) or 5 µM FCCP (Low) for 10 minutes. The fluorescent intensities for both J-aggregates and monomeric forms of JC-10 were measured with a flow cytometer using FL1 and FL2 channels. Uncompensated data (left column) were compared with compensated data (right column).
Protocols
References
This product has been referenced in:
- Khurana S et al. Antiapoptotic actions of methyl gallate on neonatal rat cardiac myocytes exposed to H2O2. Oxid Med Cell Longev 2014:657512 (2014). Flow Cyt ; Rat . Read more (PubMed: 24672637) »
- Liu Z et al. Potent Half-Sandwich Iridium(III) Anticancer Complexes Containing C(?)N-Chelated and Pyridine Ligands. Organometallics 33:5324-5333 (2014). Read more (PubMed: 25328266) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"