How does JC1 work?

How does JC1 work?

JC-1 is a lipophilic, cationic dye that can selectively enter into mitochondria and reversibly change color from green to red as the membrane potential increases. In healthy cells with high mitochondrial (ΔψM), JC-1 spontaneously forms complexes known as J-aggregates with intense red fluorescence.

What does JC-1 reveal about mitochondria?

The membrane-permeant JC-1 dye is widely used in apoptosis studies to monitor mitochondrial health. JC-1 dye can be used as an indicator of mitochondrial membrane potential in a variety of cell types, including myocytes and neurons, as well as in intact tissues and isolated mitochondria.

Does JC-1 increase mitochondrial membrane potential?

mitochondrial membrane potential in human fibroblast cells was determined by JC-1. FCCP treated cells showed increased monomeric (green) signal which supported change of mitochondrial membrane potential compared to control cells.

What is JC-1 staining?

JC-1 is a novel cationic carbocyanine dye that accumulates in mitochondria. The dye exists as a monomer at low concentrations and yields green fluorescence, similar to fluorescein. At higher concentrations, the dye forms J-aggregates that exhibit a broad excitation spectrum and an emission maximum at ∼590 nm.

What does MitoTracker stain?

MitoTracker Red CMXRos is a red-fluorescent dye that stains mitochondria in live cells and its accumulation is dependent upon membrane potential. The dye is well-retained after aldehyde fixation.

What is the mitochondrial membrane potential?

The mitochondrial membrane potential (ΔΨm) generated by proton pumps (Complexes I, III and IV) is an essential component in the process of energy storage during oxidative phosphorylation. Together with the proton gradient (ΔpH), ΔΨm forms the transmembrane potential of hydrogen ions which is harnessed to make ATP.

How does mitochondria power the cell?

Known as the “powerhouses of the cell,” mitochondria produce the energy necessary for the cell’s survival and functioning. Through a series of chemical reactions, mitochondria break down glucose into an energy molecule known as adenosine triphosphate (ATP), which is used to fuel various other cellular processes.

What is mitochondrial depolarisation?

When a mitochondrion depolarizes, it stops producing ATP, and as more mitochondria depolarize in the cell due to the increased mPTP open probability, the cytosolic ATP decreases. Since ATP is required for the SERCA pump, a low ATP level could impair the SERCA activity to promote Ca2+ alternans.

What is mitochondrial membrane potential?

How does annexin V work?

Annexin A5 (or annexin V) is a cellular protein in the annexin group. In flow cytometry, annexin V is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of apoptosis when it is on the outer leaflet of the plasma membrane.

What does MitoTracker measure?

MitoTracker green (MTG) and nonylacridine orange (NAO) are fluorescent probes which have been commonly used to assess mitochondrial mass. This is based on the assumption that both MTG and NAO accumulate in mitochondria regardless of the mitochondrial transmembrane potential (ΔΨm).

Can a MitoTracker be fixed?

For MitoTracker® Red CMXRos, we have found that fixing with 3.7% formaldehyde in complete growth medium at 37°C for 15 minutes works well for endothelial cells.

How do you make a dilution buffer for JC-1?

Prepare 1X Supplemented Dilution Buffer by adding 2 mL FBS to 18 mL 1X Dilution Buffer. 11.31 mM JC-1 Stock Solution Allow the lyophilized vial JC-1 and the DMSO to warm to room temperature. Reconstitute the JC-1 by adding 766.8 µL DMSO to the 500 µg vial of JC-1.

What is the JC1 assay kit for HepG2 cells?

JC-1 assay result in HepG2 cells treated with CCCP. JC1 – Mitochondrial Membrane Potential Assay Kit (ab113850). HepG2 cells were seeded and labeled according to section 11.2 of the protocol.

What is the mitoprobe™ JC-1 assay kit?

The MitoProbe™ JC-1 Assay Kit is a quick and reliable assay used to detect changes in mitochondria by flow cytometry. View a selection guide for all apoptosis assays for flow cytometry. Selective to mitochondrial changes The membrane-permeant JC-1 dye is widely used in apoptosis studies to monitor mitochondrial health.

What is the staining protocol for JC-1 staining?

The JC-1 staining protocol is very simple: – wash cells in dilution buffer or PBS – add JC solution – incubate for 30 min at 37ºC for suspension cells, or 10 min for adherent cells