What does DiOC6 stain?
DiOC6 staining reveals organelle structure and dynamics in living yeast cells. Cell Motil Cytoskeleton.
What is mitochondrial polarization?
Mitochondrial function is directly linked to mitochondrial polarization state. Intact mitochondria are polarized, i.e. they sustain a highly charged (negative inside) membrane potential for full functionality12.
What is mitochondrial membrane potential assay?
The Mitochondrial Membrane Potential Assay Kit (II) is a fluorescent assay that detects the mitochondrial membrane potential in living cells.
What does decreased mitochondrial membrane potential mean?
328,329 Loss of mitochondrial membrane potential is a signal of bioenergetic stress and may result in the release of apoptotic factors leading to cell death.
What is MMP assay?
The MMP assay protocol uses a fluorescence resonance energy transfer (FRET) peptide as a generic MMP activity indicator. In the intact FRET peptide, the fluorescence of one part is quenched by another. After cleavage into two separate fragments by MMPs, the fluorescence is recovered.
How do you measure mitochondrial activity?
The efficiency of oxidative phosphorylation can be measured in isolated mitochondria by high-resolution respirometry [30]. The general concept underlying this method is to measure the amount of oxygen consumed for a given amount of ATP synthesized by the mitochondria.
What causes mitochondrial depolarization?
Moreover, there is a positive feedback loop between mitochondrial ROS production and SR Ca2+ release, i.e., leaky RyRs trigger more Ca2+ release resulting in more Ca2+ sequestered in mitochondria, which can trigger the opening of mPTP, leading to mitochondria depolarizations [34,35].
What causes mitochondrial uncoupling?
Mitochondrial uncoupling can be caused by a variety of conditions and molecules that exert an influence not only on proton leak and cation cycling but also on proton slip within the proton pumps and on the structural integrity of the mitochondria.
How do I find my MMP activity?
Matrix metalloproteinases (MMP) belong to a distinguished class of zinc-dependent endopeptidases. Zymography is a semi-quantitative tool for determining the activity of different MMP isoenzymes in a variety of biological samples.
Does dioc 6 (3) change mitochondrial membrane potential?
Thus DiOC 6 (3) predominantly reported changes in mitochondrial membrane potential and not transplasmalemmal membrane potential. The apparent discordance in membrane potential results obtained using carbocyanine dye and the lipophilic cation TPMP + may be explained on the basis of differences in probe distribution.
How does dioc 6 bind to living cells?
Binding to these structures occurs via the dye’s hydrophilic groups. DiOC 6 can be used to label living cells, however they are quickly damaged due to the dye’s extreme phototoxicity, so cells stained with this dye can only be exposed to light for short periods of time. When exposed to blue light, the dye fluoresces green.
What is dioc 6 (3) used for?
Membrane potential changes have been monitored in human neutrophils using the carbocyanine dye DiOC 6 (3) (3,3′-dihexyloxacarbocyanine iodide). Fluorescence microscopy demonstrated that the carbocyanine dye DiOC 6 (3) partitions predominantly into subcellular organelles, probably mitochondria.
What is the standard temperature of DiOC6?
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).?) DiOC6 (3,3′-dihexyloxacarbocyanine iodide) is a fluorescent dye used for the staining of a cell’s endoplasmic reticulum, vesicle membranes and mitochondria.