What is h2dcfda (DCFH-da)?
DCF is a highly fluorescent compound which can be detected by fluorescence spectroscopy with excitation / emission at 495 nm / 529 nm. H2DCFDA / DCFH-DA / DCFDA assay protocol / ROS assay protocol summary (microplate): – collect suspension cells in tube / seed and allow attachment of adherent cells in 96-well plate.
What is h2dcfda – Cellular ROS assay kit?
DCFDA – Cellular ROS Assay Kit / Reactive Oxygen Species Assay Kit (ab113851) uses the cell permeant reagent 2’,7’ –dichlorofluorescin diacetate (DCFDA, also known as H2DCFDA, DCFH-DA, and DCFH) to quantitatively assess reactive oxygen species in live cell samples.
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.
Why does cm-h2 DCFDA increase fluorescence in the control cells?
I labeled my cell with CM-H 2 DCFDA for reactive oxygen detection, but upon illuminating the cell there is a significant increase in fluorescence in the control cells. Why? If the cell is overloaded with dye, the high intracellular concentration of the dye may lead to dye-dye quenching.