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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.

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based (quantitative)

Assay time

40m

Images

What's included?

300 Test
Components
10X Dilution Buffer (Sterile)
1 x 10 mL
20 mM DCFDA (Label)
1 x 35 µL
55mM TBHP
1 x 50 µL

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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.

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based (quantitative)

Assay time

40m

Assay Platform

Microplate reader, Fluor. microscope, Flow cyt.

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

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.

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

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.

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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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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