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DCFDA Assay Kit ab113851 includes DCFDA reagent, buffer, TBHP positive control, and a full protocol, for a fast way to set up a DCFDA assay on live cells. DCFDA taken up by cells is oxidized by reactive oxygen species with fluorescent readout (Ex/Em 485/535 nm) by microscopy, flow cytometer or plate reader.
Is this your first time purchasing this assay kit? Get 20% off one kit by entering code 20OFF-T5A8Q at checkout. T&C’s apply – click here. Offer only valid when purchasing directly from Abcam in Europe, North America, Australia, and Singapore.
Fluorescent
Suspension cells, Adherent cells
Cell-based (quantitative)
40m
DCFDA Assay Kit ab113851 includes DCFDA reagent, buffer, TBHP positive control, and a full protocol, for a fast way to set up a DCFDA assay on live cells. DCFDA taken up by cells is oxidized by reactive oxygen species with fluorescent readout (Ex/Em 485/535 nm) by microscopy, flow cytometer or plate reader.
Is this your first time purchasing this assay kit? Get 20% off one kit by entering code 20OFF-T5A8Q at checkout. T&C’s apply – click here. Offer only valid when purchasing directly from Abcam in Europe, North America, Australia, and Singapore.
Fluorescent
Suspension cells, Adherent cells
Cell-based (quantitative)
40m
Microplate reader, Fluor. microscope, Flow cyt.
Blue Ice
-20°C
-20°C
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 semi quantitatively assess reactive oxygen species in live cell samples.
DCFDA / H2DCFDA / DCFH-DA / DCFH is a fluorogenic dye that measures hydroxyl, peroxyl and other reactive oxygen species (ROS) activity within the cell. NB: DCFDA and DCFA are also available as free molecules as ab145286 (Carboxy-DCFDA N-succinimidyl ester) and ab145439 (5(6)-Carboxy-2',7'-dichlorofluorescein).
The DCFDA assay protocol is based on the diffusion of DCFDA / H2DCFDA / DCFH-DA / DCFH into the cell. It is then deacetylated by cellular esterases to a non-fluorescent compound, which is later oxidized by ROS into 2’, 7’ –dichlorofluorescein (DCF). DCF is highly fluorescent and is detected by fluorescence spectroscopy with excitation / emission at 485 nm / 535 nm.
DCFDA assay protocol / ROS assay protocol summary (microplate):
- collect suspension cells in tube / seed and allow attachment of adherent cells in 96-well plate
- wash in buffer
- stain with DCFDA for 30 min (suspension) / 45 min (adherent), wash with buffer
- if suspension cells, transfer to microplate
- analyze with microplate reader
DCFDA assay protocol / ROS assay protocol summary (flow cytometry):
- collect cells in tubes
- stain with DCFDA for 30 min (without washing)
- analyze with flow cytometer
DCFDA assay protocol / ROS assay protocol summary (fluorescent microscopy):
- wash adherent cells with buffer
- stain with DCFDA for 45 min
- wash in buffer
- analyze with fluorescent microscope
- maintain low light conditions to reduce photo-bleaching
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