MitoPT TMRM Mitochondrial Depolarization Assay Kit | MitoPT TMRM assay kit

MitoPT TMRM Mitochondrial Depolarization Assay Kit

Cat. Num. AAA258008

• Synonyms: MitoPT TMRM Mitochondrial Depolarization; MitoPT TMRM Mitochondrial Depolarization Assay Kit; MitoPT TMRM assay kit

• Reagents: TMRM
• Sample Protocol: Culture cells up to 1 x 10^6 cells/mL. Expose cells to apoptosis inducing agent or metabolic stress environment using your experimental protocol. Reconstitute the TMRM reagent with 100 uL DMSO to form the stock concentrate (may be frozen for future use). Dilute the stock concentrate with 1X PBS, cell culture media, or wash buffer to achieve a 10 uM intermediate stock for spiking cell aliquots. Spike cell cultures with 10 uM TMRM to achieve a 20-200 nM TMRM concentration. Incubate cells in TMRM (20-200 nM) solution, 15-20 minutes at 37 degree C in dark. Wash cells once in wash buffer if using > 50 nM TMRM staining concentration. Analyze data using a fluorescence microscope, plate reader, or flow cytometer.
• Target: Mitochondrial Depolarization
• Excitation / Emission: 548 nm / 573 nm
• Method of Analysis: Flow Cytometer, Fluorescence Microscope, Fluorescence Plate Reader
• Types of Samples: Cell Culture, Tissue
• Preparation and Storage: Mixed Storage Conditions: CCCP at -20 degree C; Other Kit Components at 2 - 8 degree C

Typical Testing Data/Standard Curve (for reference only)

Related Product Information for MitoPT TMRM assay kit

Description: MitoPT TMRM assay kits utilize the well-characterized, lipophilic but slightly cationic fluorescent dye TMRM to detect mitochondrial membrane permeability. Use of the MitoPT TMRM kit allows for the easy distinction of apoptotic or mitochondrial membrane-compromised cells from non-apoptotic cells with polarized mitochondria. Cells with polarized mitochondria will fluoresce bright red (em max: 573 nm); cells with membrane- compromised cells will exhibit a dimished fluorescence or no signal. The MitoPT TMRM mitochondrial membrane permeability assay can be used in conjunction with existing apoptosis or metabolic stress protocols. Each kit includes the TMRM potentiometric dye reagent, a 10X wash buffer for removing any excess TMRM dye after staining, and the generic mitochondrial membrane depolarizer, carbonylcyanide m- chlorophenylhydrazone (CCCP). TMRM-stained cells can be analyzed using flow cytometry, fluorescence microscopy, and fluorescence plate reader evaluation methodology.

Background: Detect Mitochondrial Permeability Transition and Mitochondrial Membrane Depolarization with the MitoPT TMRM Assay ICT's MitoPT TMRM Assay utilizes the well-characterized potentiometric fluorescent dye tetramethylrhodamine methyl ester (TMRM) to detect mitochondrial permeability and membrane depolarization in whole, living cells. The rhodamine-based TMRE and TMRM dyes offered in ICT's MitoPT cell-based assay kits are categorized as slow membrane redistribution dyes. The lipophilic structures of hese dyes allow them to easily penetrate cell and mitochondrial lipid bilayer membrane barriers. Their potentiometric fluorescent properties allow for the detection of the mitochondrial permeability transition in whole cells via fluorescence analysis methods.

Product Categories/Family for MitoPT TMRM assay kit

Apoptosis Assays/Apoptosis Detection

References

Plasek, J. and K. Sigler. 1996. Slow fluorescent indicators of membrane potential: a survey of different approaches to probe response analysis. J. Photoch. Photobio. B. 33: 101-124. Ehrenberg, B., V. Montana, M. Wei, J.P. Wuskell, and L.M. Loew. 1988. Membrane potential can be determined in individual cells from the Nernstian distribution of cationic dyes. Biophys. J. 53: 785-794. Waggoner, A.S. 1979. Dye indicators or membrane potential. Ann Rev. Biophys Bioeng. 8: 47-68. o, R.C. and L.W. Grotyohann. 1999. Measurement of mitochondrial membrane potential using fluorescent rhodamine derivatives. Biophys. J. 76: 469-477. Wlodkowic, D., J. Skommer, and J. Pelkonen. 2006. Multiparametric analysis of HA14-1 induced apoptosis in follicular lymphoma cells. Leukemia Res. 30: 1187-1192.