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 (ICT943) 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.
Jurkat T cells were treated with DMSO or 1 μM staurosporine as shown followed by staining with Mitochondrial Permeability Transition: MitoPT JC-1 Kit (ICT943) 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.
Jurkat cells were stained with MitoPTTM JC-1 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)
The MitoPT™ JC-1 kit uses a quick and easy staining method to clearly differentiate between non-apoptotic and apoptotic cells through mitochondrial functionality.
Approx. Protein Concentrations
Reagents In The Kit
MitoPTTM JC-1 Reagent - lyophilized 10X Assay Buffer, 60mL 125uL vial 50mM Carbonylcyanide m - chlorophenylhydrazone (CCCP) concentrate in DMSO.
Preparing The Antibody
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 MitoPTTM JC-1 kit.
The MitoPTTM 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.
Store the unopened kit (and each unopened component) at -20oC until the expiration date. Protect the MitoPTTM JC-1 reagent from light at all times.
Once reconstituted, the 100X MitoPTTMJC-1 stock should be stored at -20oC protected from light.
Please see label for expiry date.
MitoPTTM is a trademark of Immunochemistry Technologies, LLC.
For research purposes only
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.
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.