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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.

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based (quantitative)

Assay time

40m

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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 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.

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 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

Product promise

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Product protocols

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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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