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Chemical Structure| 4091-99-0 Chemical Structure| 4091-99-0

Structure of H2DCFDA
CAS No.: 4091-99-0

Chemical Structure| 4091-99-0

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H2DCFDA (DCFH-DA) is a cell-permeable probe used for detecting intracellular reactive oxygen species (ROS) with excitation/emission at 488/525 nm.

Synonyms: DCFH-DA; 2',7'-Dichlorodihydrofluorescein diacetate; DCFH

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Product Citations

Product Citations

Wang, Lei ; Li, Zhong-Yao ; Zhong, Chong-Lei ; Teng, Zi-Yang ; Wang, Bin ; Rehman, Asma , et al.

Abstract: Background: Sepsis is a life-threatening infectious disease characterized by an uncontrolled infammatory response and consequent multi-organ dysfunction. The kidneys, as primary excretory organs with high blood fow, are particularly susceptible to damage during sepsis. Nonetheless, the existing treatment options for sepsis-associated acute kidney injury (SA-AKI) are still restricted. Nanomedicine, especially carbon dots (CDs), has attracted considerable interest lately for outstanding biomedical characteristics. Methods: To avoid the generation of toxic efects, the natural CDs derived from Ziziphi Spinosae Semen (Z-CDs) were synthesized employing a hydrothermal method. The free radical scavenging capabilities of Z-CDs were evaluated by utilizing ABTS assay, NBT method, and Fenton reaction. A lipopolysaccharide (LPS)-stimulated RAW 264.7 cell model was used to explore the therapeutic potential of Z-CDs on cellular oxidative stress and infammation. The CuSO4-induced zebrafsh infammation model and LPS-exposed SA-AKI mouse model were employed to assess the therapeutic efcacy of Z-CDs in vivo. Results: The synthesized Z-CDs exhibited distinctive unsaturated surface functional groups, which confer exceptional biocompatibility and the ability to scavenge free radicals. Moreover, Z-CDs were particularly efective in eliminating excess reactive oxygen species (ROS) in cells, thus protecting mitochondrial function from oxidative damage. Notably, Z-CDs have demonstrated signifcant therapeutic benefts in protecting kidney tissue in SA-AKI mouse model with minimizing side efects. In mechanism, Z-CDs efectively reduced ROS production, thereby alleviating infammatory responses in macrophages through the suppression of the NF-κB pathway. Conclusions: This study developed a multifunctional nanomedicine derived from traditional medicinal herb, providing a promising pathway for the advancement of innovative drug therapies to treat SA-AKI.

Keywords: Sepsis ; Acute kidney injury ; Carbon dots ; Phytochemicals ; Antioxidant ; NF-κB pathway

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Product Details of H2DCFDA

CAS No. :4091-99-0
Formula : C24H16Cl2O7
M.W : 487.29
SMILES Code : O=C(O)C1=CC=CC=C1C2C3=C(OC4=C2C=C(Cl)C(OC(C)=O)=C4)C=C(OC(C)=O)C(Cl)=C3
Synonyms :
DCFH-DA; 2',7'-Dichlorodihydrofluorescein diacetate; DCFH
MDL No. :MFCD00128955
InChI Key :PXEZTIWVRVSYOK-UHFFFAOYSA-N
Pubchem ID :77718

Safety of H2DCFDA

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H315-H319-H335
Precautionary Statements:P261-P305+P351+P338

Isoform Comparison

Biological Activity

Description
H2DCFDA (DCFH-DA) is a probe permeable to cells and is employed for the detection of intracellular reactive oxygen species (ROS), with an excitation and emission wavelength of 488/525 nm respectively[1].

In Vitro:

Cell Line
Concentration Treated Time Description References
BMDMs 1 µM Used to measure intracellular ROS levels PMC9774280
Human primary keratinocytes (NHEK) 10 μM 30 s To detect H2O2 uptake after AQP3 knockdown, results showed that H2O2 uptake was significantly reduced in AQP3 knockdown cells PMC5628617
Mouse primary keratinocytes 10–300 μM 15 s To detect AQP3-mediated H2O2 transport, results showed that H2O2 uptake was significantly higher in WT cells compared to AQP3−/− cells PMC5628617
SH-SY5Y cells 5 μM 10 min To detect intracellular levels of hydrogen peroxide as a marker of reactive oxygen species (ROS), the results showed that the H2DCFDA probe effectively detected intracellular ROS levels. PMC7985515
HMSCs 1 μM To measure ROS levels in SIRT7+/+ and SIRT7−/− hMSCs, the results showed that SIRT7 deficiency led to increased ROS levels. PMC7305295
NHDF cells 25 µM 30 min To detect PM2.5-induced intracellular ROS levels, results showed that ginsenoside Rb1 pretreatment significantly reduced PM2.5-induced ROS production. PMC6769862
HaCaT cells 25 µM 30 min To detect PM2.5-induced intracellular ROS levels, results showed that ginsenoside Rb1 pretreatment significantly reduced PM2.5-induced ROS production. PMC6769862
SC SOD1G93A astrocytes 5 µM To measure ROS levels in SC SOD1G93A astrocytes, results showed increased H2DCFDA fluorescence intensity, indicating elevated ROS content. PMC8472526
MC SOD1G93A astrocytes 5 µM To measure ROS levels in MC SOD1G93A astrocytes, results showed increased H2DCFDA fluorescence intensity, indicating elevated ROS content. PMC8472526
Mouse Embryonic Fibroblasts (MEFs) 3 µM 45 min To assess H2O2 levels in Redd1/C0//C0 cells and tissues, showing significantly lower H2O2 levels in Redd1/C0//C0 cells and tissues compared to wild-type controls PMC4421852
Mouse peritoneal macrophages 2 µM 30 min To measure ROS levels in macrophages after bacterial phagocytosis PMC6861186

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
Mice IL-23-induced psoriasis model Intradermal injection into the ear 500 ng Daily for 4 days To investigate the role of AQP3 in psoriasis development, results showed that psoriasis-like inflammation was significantly reduced in AQP3?/? mice PMC5628617

Protocol

Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

2.05mL

0.41mL

0.21mL

10.26mL

2.05mL

1.03mL

20.52mL

4.10mL

2.05mL

Dissolving Methods
Please choose the appropriate dissolution scheme according to your animal administration guide.For the following dissolution schemes, clear stock solution should be prepared according to in vitro experiments, and then cosolvent should be added in turn:

in order to ensure the reliability of the experimental results, the clarified stock solution can be properly preserved according to the storage conditions; The working fluid for in vivo experiment is recommended to be prepared now and used on the same day;

The percentage shown in front of the following solvent refers to the volume ratio of the solvent in the final solution; If precipitation or precipitation occurs in the preparation process, it can be assisted by heating and/or ultrasound.
Protocol 1
Protocol 2

References

 

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