DCFDA / H2DCFDA - Cellular ROS Assay Kit

(HB7375)
Technical documents: Datasheet

Product overview

Name DCFDA / H2DCFDA - Cellular ROS Assay Kit
Biological description

DCFDA / H2DCFDA is a cell permeable fluorescent probe that is redox sensitive and used to measure the concentration of reactive oxygen species (ROS) within a population of cells. DCFDA / H2DCFDA diffuses into cells where it is hydrolysed by intracellular esterases into a non-fluorescent and non-cell permeable intermediate. Upon reaction with ROS this forms the fluorescent compound 2’,7’ –dichlorofluorescein (DCF) which is excited at 485nm and emits at 535nm. Pyocyanin is included within this kit as a positive control. Pyocyanin promotes the formation of ROS through inactivation of catalase and depleting reduced glutathione.

 

This kit contains:

  • DCFDA / H2DCFDA assay reagent
  • DMSO
  • Lyophilised Pyocyanin
  • 10x assay buffer
Description

Kit for measurement of reactive oxygen species (ROS) within cells.

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Images

Figure 1. Concentration response of pyocyanin in HEK293T cells using HB7375 DCFDA / H2DCFDA - Cellular ROS Assay Kit

Pyocyanin induces oxidative stress in HEK293T cells which is detected as an increase in fluorescence using the H2DCFDA probe. Method: HEK293T cells were seeded at 50,000 cells per well in a 96 well microplate before being cultured overnight. The following day cells were washed with 1x assay buffer before being incubated in 20µM H2DCFDA for 45 minutes at 37°C. Cells were then washed again with 1x assay buffer before being incubated in differing concentrations of pyocyanin in assay buffer for 1 hour at 37°C. Fluorescence was then read in a microplate reader (excitation at 485nm and emission at 535nm).

Optical Data

Emission color Green
Max excitation wavelength 485nm
Max emission wavelength 535nm
Closest laser lines 488 nm
Cell permeable Yes

Biological Data

Application notes

Please follow this link to a full DCFDA / H2DCFDA - Cellular ROS Assay Kit protocol

Solubility & Handling

Storage instructions

-20°C

Important This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use

Calculators

Molarity

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More Info

Dilution

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Chemical Data

Chemical name

2-[3,6-Bis(acetyloxy)-2,7-dichloro-9H-xanthen-9-yl]benzoic acid

Molecular Weight 485.27
Chemical structure H2DCFDA [4091-99-0] Chemical Structure
Molecular Formula

C24H14CI2O7

CAS Number 4091-99-0
PubChem identifier 77718
SMILES CC(=O)OC1=C(C=C2C(C3=CC(=C(C=C3OC2=C1)OC(=O)C)Cl)C4=CC=CC=C4C(=O)O)Cl
InChiKey PXEZTIWVRVSYOK-UHFFFAOYSA-N

References for DCFDA / H2DCFDA - Cellular ROS Assay Kit

References are publications that support the biological activity of the product
  • Detection of Total Reactive Oxygen Species in Adherent Cells by 2',7'-Dichlorodihydrofluorescein Diacetate Staining.

    Kim H et al (2020) Journal of visualized experiments : JoVE :
  • The involvement of TLR2 in cytokine and reactive oxygen species (ROS) production by PBMCs in response to Leishmania major phosphoglycans (PGs).

    Kavoosi G et al (2009) Parasitology 136 : 1193-9

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