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.

How does jc1 work?

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 a JC-1 assay?

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 CCCP treatment?

A classic oxidative phosphorylation uncoupler, carbonyl cyanide m-chlorophenylhydrazone (CCCP), is frequently used to induce the opening of the permeability transition pore on the mitochondrial membrane, leading to the dissipation of mitochondrial membrane potential (ΔΨm)8.

What is FCCP in mitochondria?

FCCP is a potent uncoupler of oxidative phosphorylation in mitochondria that disrupts ATP synthesis by transporting protons across cell membranes.

What does MitoTracker deep red measure?

First, MitoTracker Deep Red is actually a mitochondrial potential-dependent dye though this is usually not highlighted in the manufacturer’s manual. It has been used as an index for mitochondrial potential (Lugli et al., 2005; Zhou et al., 2011; Greene et al., 2012).

What does MitoTracker red do?

MitoTracker® Red FM is a far red-fluorescent dye (abs/em ∼581/644 nm) that stains mitochondria in live cells and its accumulation is dependent upon membrane potential. For Research Use Only. Not for use in diagnostic procedures.

Is CCCP toxic?

Causes skin irritation. Material may be irritating to the mucous membranes and upper respiratory tract. May cause respiratory system irritation. Toxic if inhaled, swallowed, or in contact with skin.

What is the difference between FCCP and CCCP?

FCCP applied for 30 minutes at 10 microM reduces the number of moving organelles by 78% and a similar treatment with CCCP causes a reduction of 47%. At 100 microM either compound abolishes all directed movements both in neurites and in cultured 3T3 cells.

Why is FCCP used?

FCCP is an protonophore which is widely used to investigate the role of mitochondria in cellular function. FCCP is a potent uncoupler of mitochondrial oxidative phosphorylation. FCCP disrupts ATP synthesis by transporting protons across the mitochondrial inner membrane, interfering with the proton gradient.

Why does FCCP increase oxygen consumption?

Maximal respiration: The maximal oxygen consumption rate attained by adding the uncoupler FCCP. FCCP mimics a physiological “energy demand” by stimulating the respiratory chain to operate at maximum capacity, which causes rapid oxidation of substrates (sugars, fats, and amino acids) to meet this metabolic challenge.

What diseases can flow cytometry detect?

Flow cytometry is a laser-based technique used to detect and analyze the chemical and physical characteristics of cells or particles….Results and Follow-Up

• Multiple myeloma.
• Non-Hodgkin lymphomas (both B-cell and T-cell).
• Acute myeloid leukemia.
• Acute lymphoblastic leukemia (ALL).
• Chronic lymphocytic leukemia (CLL).

What is considered positive in flow cytometry?

We can set a marker on the control sample and everything that appears to the right in the test sample is deemed positive. Figure 2. Negative sample (A) and test sample (B).

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. 2.3 Rinsing the cells. After fixation, rinse the cells several times in buffer.

What does MitoTracker green 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).

How does MitoTracker green work?

MitoTracker Green (MTG) is a mitochondrial-selective fluorescent label commonly used in confocal microscopy and flow cytometry. It is expected that this dye selectively accumulates in the mitochondrial matrix where it covalently binds to mitochondrial proteins by reacting with free thiol groups of cysteine residues.

What is CCCP used for?

CCCP is a protonophore which is a widely used uncoupler of mitochondrial oxidative phosphorylation. CCCP disrupts ATP synthesis by transporting protons across the mitochondrial inner membrane, interfering with the proton gradient.

What does CCCP treatment do?

The CCCP treatment was found to significantly reduce ATP content, increase the amount of phosphorylated AMP-activated protein kinase and elevate reactive oxygen species levels in oocytes.