Jurkat cells were cultured in the absence (healthy) or presence (dying) of staurosporine to induce apoptosis and then stained with TMRM dye. Cells were analyzed using a Nikon Eclipse E800 photomicroscope using a green excitation filter (510-560 nm) in tandem with a 570-620 nm emission filter. Apoptotic cells with depolarized mitochondria exhibit reduced orange/red fluorescence
The MitoPTTM TMRM 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 TMRM Reagent - lyophilized - to make up 1mM stock 10X Assay Buffer, 125mL 600uL 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 TMRM (tetramethylrhodamine methyl ester), that has been incorporated into the MitoPTTM TMRM kit.
The MitoPTTM TMRM reagent easily penetrates cells and enters the mitochondria. It aggregates in the mitochondria of non-apoptotic cells and fluoresces bright orange/red, whilst in apoptotic cells it diffuses throughout the cell. Once dispersed, the reagent assumes a monomeric form and exhibits a reduced orange/red fluorescence level. This allows an easy distinction between apoptotic and non-apoptotic 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 TMRM reagent from light at all times.
Once reconstituted, the 1mM MitoPTTM TMRM 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.