Description
Store kit at 4°C in the dark immediately upon receipt.
TMRE-Mitochondrial Membrane Potential Assay Kit ab113852 is used for quantifying changes in mitochondrial membrane potential in live cells by flow cytometry, microplate spectrophotometry and fluorescent microscopy.
TMRE (tetramethylrhodamine, ethyl ester) is used to label active mitochondria. TMRE is a cell permeant, positively-charged, red-orange dye that readily accumulates in active mitochondria due to their relative negative charge. Depolarized or inactive mitochondria have decreased membrane potential and fail to sequester TMRE. NB: TMRE is also available as free molecule as ab274305 (Tetramethylrhodamine ethyl ester).
The TMRE protocol also uses FCCP (carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone), which is a ionophore uncoupler of oxidative phosphorylation. Treating cells with FCCP eliminates mitochondrial membrane potential and TMRE staining. TMRE is suitable for the labeling of mitochondria in live cells and is not compatible with fixation.
TMRE protocol summary:
– add FCCP to appropriate control cell samples and incubate for 10 min
– incubate with TMRE for 15-30 min, pellet (suspension cells) / remove media (adherent cells) and wash with PBS / 0.2% BSA
– analyze with micro-plate reader at Ex/Em 549/575 nm, flow cytometer using 488nm laser for excitation and at emission 575 nm, or fluorescent microscope.
TMRE is only suitable for use with live (not fixed) cells. **Related assays** Review the to learn about kits to perform a , and . Review the to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader. **How other researchers have used TMRE assay kit ab113852** Maiti AK et al. of the University of Gothenburg, Sweden, used TMRE assay kit ab113852 to identify that C. rodentium infection in mice* reduced mitochondrial transmembrane potential. In a subsequent paper, they identified that this effect was reduced by treatment with vasoactive intestinal peptide (VIP). *a model for enteropathogenic E. coli