How does CM h2dcfda work?
CM-H2DCFDA passively diffuses into cells, where its acetate groups are cleaved by intracellular esterases and its thiol-reactive chloromethyl group reacts with intracellular glutathione and other thiols.
Do mitochondria generate ROS?
Mitochondria are an important source of ROS (reactive oxygen species) within most mammalian cells [1–8]. This ROS production contributes to mitochondrial damage in a range of pathologies and is also important in redox signalling from the organelle to the rest of the cell [3,9].
Are mitochondrial diseases genetic?
Mitochondrial diseases are long-term, genetic, often inherited disorders that occur when mitochondria fail to produce enough energy for the body to function properly. One in 5,000 individuals has a genetic mitochondrial disease. Symptoms, diagnosis and treatment are discussed.
What is h2dcf DA?
The cell-permeant 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) (also known as dichlorofluores cin diacetate) is a chemically reduced form of fluorescein used as an indicator for reactive oxygen species (ROS) in cells, for example to detect the generation of reactive oxygen intermediates in neutrophils and …
What is the difference between superoxide and peroxide?
The primary difference between a peroxide and a superoxide lies in the oxidation state of the oxygen atom. In a peroxide, the oxidation state of oxygen is known to be -1 whereas in a superoxide, the oxidation state of oxygen is known to be -1/2.
What does h2dcfda stand for?
The cell-permeant 2′,7′-dichlorodihydrofluorescein diacetate (H 2 DCFDA) (also known as dichlorofluores cin diacetate) is a chemically reduced form of fluorescein used as an indicator for reactive oxygen species (ROS) in cells, for example to detect the generation of reactive oxygen intermediates in neutrophils and macrophages.
What is H2 DCFDA assay?
In the presence of ROS, predominantly H 2 O 2, H 2 DCF is rapidly oxidized to 2′,7′-dichlorofluorescein (DCF), which is highly fluorescent, with excitation and emission wavelengths of 498 and 522 nm, respectively. The H 2 DCFDA assay can provide reliable measurements of H 2 O 2 levels in cells and mitochondrial preparations.
What is the cell permeability of cm-h2 DCFDA?
CM-H 2 DCFDA is cell permeable and becomes fluorescent when the dihydrofluorescein is oxidized to fluorescein by intracellular ROS. 2.
Why is my h2dcfda dye not working?
Some dyes, such as H2DCFDA, require esterase cleavage, so don’t incubate in the presence of serum (which contains esterases that can prematurely cleave the dye). If your positive control does not show significant change compared to the negative control, try increasing the concentration and label time for the dye.