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Mitochondrial Permeability Transition:MitoPT™ JC-1 Kit

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Mito Permeability Transition Kit - 400 Tests thumbnail image 1 Mito Permeability Transition Kit - 400 Tests thumbnail image 2 Mito Permeability Transition Kit - 400 Tests thumbnail image 3 Mito Permeability Transition Kit - 400 Tests thumbnail image 4
Mito Permeability Transition Kit - 400 Tests gallery image 1
Jurkat T cells were stained with MitoPT JC-1 (ICT944) and viewed through a fluorescence microscope using a broad band path filter. Non-apoptotic cells exhibit red stained mitochondria (2 cells at the top). Apoptotic cells at varying stages of mitochondrial membrane potential appear green (3 cells at the bottom).
Mito Permeability Transition Kit - 400 Tests gallery image 2
Jurkat T cells were treated with DMSO or 50 μM CCCP for 1 hour followed by staining with Mitochondrial Permeability Transition: MitoPT JC-1 Kit (ICT944) for 15-30 minutes. Loss of mitochondrial potential results in a reduction in red fluorescence and an increase in green fluorescence. Data acquired on the ZE5 Cell Analyzer.
Mito Permeability Transition Kit - 400 Tests gallery image 3
Jurkat T cells were treated with DMSO or 1 μM staurosporine as shown followed by staining with Mitochondrial Permeability Transition: MitoPT JC-1 Kit (ICT944) for 15-30 minutes. Loss of mitochondrial potential results in a reduction in red fluorescence and an increase in green fluorescence. Data acquired on the ZE5 Cell Analyzer.
Mito Permeability Transition Kit - 400 Tests gallery image 4
Published customer image:
The MitoPT JC-1 kit (ICT944) used to detect mitochondrial depolarization in mouse cells by immunofluorescence.
Image caption:
Polarization of mitochondrial membrane in cancer and normal cells upon UM0112176 treatment. The more yellow appear the mitochondria the higher is the ratio of JC-1 aggregates (red, strong polarization) to monomers of the dye (green, depolarized). Bar = 10μM.

From: Gizak A, Wiśniewski J, Heron P, Mamczur P, Sygusch J, Rakus D.
Targeting a moonlighting function of aldolase induces apoptosis in cancer cells.
Cell Death Dis. 2019 Sep 26;10(10):712.
doi: 10.1038/s41419-019-1968-4.
This image is from an open access article distributed under the terms of the Published customer image:
The MitoPT™ JC-1 kit (ICT943) used to detect mitochondrial depolarization in mouse cells by immunofluorescence.
Image caption:
Polarization of mitochondrial membrane in cancer and normal cells upon UM0112176 treatment. The more yellow appear the mitochondria the higher is the ratio of JC-1 aggregates (red, strong polarization) to monomers of the dye (green, depolarized). Bar = 10μM.

From: Gizak A, Wiśniewski J, Heron P, Mamczur P, Sygusch J, Rakus D.
Targeting a moonlighting function of aldolase induces apoptosis in cancer cells.
Cell Death Dis. 2019 Sep 26;10(10):712.
doi: 10.1038/s41419-019-1968-4.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.

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Mitochondrial Permeability Transition:MitoPT™ JC-1 Kit

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The MitoPT JC-1 kit uses a quick and easy staining method to clearly differentiate between non-apoptotic and apoptotic cells through mitochondrial functionality.

Reagents in the Kit
MitoPT JC-1 Reagent, 400 tests
Two bottles 10x Assay Buffer, 125 mL
CCCP (50 mM in DMSO), 600 ul.
Test Principle
An early indication of apoptosis involves a collapse in the electrochemical gradient across the mitochondrial membrane. Loss of mitochondrial membrane potential can be detected by a unique fluorescent cationic dye known as JC-1 (5,5’,6,6’-tetrachloro-1,1’,3,3’-tetraethylbenzamidazolocarbocyanin iodide) that has been incorporated into the MitoPT JC-1 kit.

The MitoPT JC-1 reagent easily penetrates cells and enters the mitochondria. It aggregates in the mitochondria of non-apoptotic cells and fluoresces red, whilst in apoptotic cells it diffuses throughout the cell. Once dispersed, the reagent assumes a monomeric form and fluoresces green. This allows an easy distinction between non-apoptotic red fluorescent cells and apoptotic green fluorescent cells which can be read with a flow cytometer, fluorescence microscope or a fluorescence plate reader using black microtitre plates.
Regulatory
For research purposes only.
Guarantee
Guaranteed until date of expiry. Please see product label.
Acknowledgements
MitoPT is a trademark of Immunochemistry Technologies, LLC.

Store the unopened kit and each unopened component at -20°C until the expiration date. Once opened, some components may be stored at 2-8°C until the expiration date. CCCP should be stored frozen. Once reconstituted with DMSO, dilute and use MitoPT reagent immediately, or store at ≤-20°C for 12 months protected from light and thawed no more than twice.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Flow Cytometry
Immunofluorescence
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Instructions For Use
Instructions for use can be found at www.bio-rad-antibodies.com/uploads/IFU/ICT944.pdf

References for Mitochondrial Permeability Transition:MitoPT™ JC-1 Kit

  1. Gizak, A. et al. (2019) Targeting a moonlighting function of aldolase induces apoptosis in cancer cells.
    Cell Death Dis. 10 (10): 712.
  2. Jasek, E. et al. (2012) Effect of histone deacetylase inhibitors trichostatin A and valproic acid on etoposide-induced apoptosis in leukemia cells.
    Anticancer Res. 32 (7): 2791-9.
  3. Xiaojuan, W. et al. (2014) In Vitro Comparative Effect of Three Novel Borate Bioglasses on the Behaviors of Osteoblastic MC3T3-E1 Cells
    Journal of Materials Science & Technology. 30 (10): 979-83.
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