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D6883

2′,7′-Dichlorodihydrofluorescein diacetate

≥97% (HPLC), powder, cell-permeable non-fluorescent probe

Synonym(s):

2′,7′-Dichlorofluorescin diacetate, DCFH-DA, 2′,7′-Dichlorodihydrofluorescein diacetate

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About This Item

Empirical Formula (Hill Notation):
C24H16Cl2O7
CAS Number:
4091-99-0
Molecular Weight:
487.29
NACRES:
NA.77
PubChem Substance ID:
24894058
UNSPSC Code:
12352200
MDL number:
MFCD00128955
Beilstein/REAXYS Number:
4610273
Assay:
≥97%
Form:
powder
Quality level:
200
Pricing and availability is not currently available.
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Product Name

2′,7′-Dichlorodihydrofluorescein diacetate, ≥97%

Quality Level

200

assay

≥97%

form

powder

storage temp.

−20°C

SMILES string

CC(=O)Oc1cc2Oc3cc(OC(C)=O)c(Cl)cc3C(c2cc1Cl)c4ccccc4C(O)=O

InChI

1S/C24H16Cl2O7/c1-11(27)31-21-9-19-15(7-17(21)25)23(13-5-3-4-6-14(13)24(29)30)16-8-18(26)22(32-12(2)28)10-20(16)33-19/h3-10,23H,1-2H3,(H,29,30)

InChI key

PXEZTIWVRVSYOK-UHFFFAOYSA-N

General description

2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) is an oxidation-sensitive fluorescent dye used to detect the intracellular reactive oxygen species (ROS) level. DCFH-DA is a cell-permeable non-fluorescent probe, that can be hydrolyzed by intracellular esterase to DCFH, which forms fluorescent DCF when oxidized by the intracellular ROS.[1]
The excitation wavelength is 504 nm and emission wavelength is 529 nm in ethanol. The use of DMSO as the solvent has not been determined, however various publications suggest that it is suitable.

Application

2′,7′-Dichlorofluorescin diacetate is suitable for use in:
  • sensitive and rapid quantitation of oxygen-reactive species in response to oxidative metabolism[2][1][3]
  • microplate assay for detecting oxidative products in phagocytic cells
  • quantitative multiwell myeloid differentiation assay

Applications include sensitive and rapid quantitation of oxygen-reactive species in response to oxidative metabolism; microplate assay for detecting oxidative products in phagocytic cells, and quantitative multiwell myeloid differentiation assay.

Preparation Note

This product should be stored in powdered format at -20°C.

Other Notes

2024 CiteAb Award Winner for Supplier Succeeding in Parkinson′s Research

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This Item
287810D6665410217
2′,7′-Dichlorodihydrofluorescein diacetate ≥97%

Sigma-Aldrich

D6883

2′,7′-Dichlorodihydrofluorescein diacetate

2′,7′-Dichlorofluorescin Diacetate Cell-permeable fluorogenic probe that is useful for the detection of reactive oxygen species (ROS) and nitric oxide (•NO) and for the determination of the degree of overall oxidative stress.

Sigma-Aldrich

287810

2′,7′-Dichlorofluorescin Diacetate

2′,7′-Dichlorofluorescein suitable for use as an indicator in chloride titration, ~90% (TLC), powder

Sigma-Aldrich

D6665

2′,7′-Dichlorofluorescein

2′,7′-Dichlorofluorescein ACS reagent

Sigma-Aldrich

410217

2′,7′-Dichlorofluorescein

form

powder

form

crystalline solid

form

powder

form

powder

assay

≥97%

assay

≥94% (HPLC)

assay

~90% (TLC)

assay

-

Quality Level

200

Quality Level

100

Quality Level

200

Quality Level

200

storage temp.

−20°C

storage temp.

−20°C

storage temp.

room temp

storage temp.

room temp

Storage Class

11 - Combustible Solids

wgk

WGK 3

ppe

Eyeshields, Gloves, type N95 (US)

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Certificates of Analysis (COA)

Lot/Batch Number
0000540774
0000533048
0000533048
0000540774
0000438975

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Katia Cortese et al.
Life sciences, 232, 116610-116610 (2019-06-30)
The aim of this study was the characterization of the in vitro cytotoxic properties of a recently isolated diterpene compound, 7β-acetoxy-20-hydroxy-19,20-epoxyroyleanone (compound 1), extracted from Salvia corrugata, versus human cell lines. We used as model study immortalized breast epithelial cells
Yuan Ji et al.
Journal of agricultural and food chemistry, 67(32), 9060-9069 (2019-07-25)
Glutathione S-transferases (GSTs) play an active role in the development of drug resistance by numerous cancer cells, including melanoma cells, which is a major cause of chemotherapy failure. As part of our continuous effort to explore why dietary polyphenols bearing
Pornthip Chaichompoo et al.
Journal of clinical pathology, 72(8), 520-524 (2019-04-24)
Iron overload is a major factor contributing to the overall pathology of thalassaemia, which is primarily mediated by ineffective erythropoiesis and shorter mature red blood cell (RBC) survival. Iron accumulation in RBCs generates reactive oxygen species (ROS) that cause cellular
View All Related Papers

Global Trade Item Number

SKUGTIN
D6883-50MG04061835361229
D6883-250MG04061838445964

Questions

1–10 of 14 Questions  

  1. Can you suggest a protocol for detection of ROS in colon tissue from rat?

    1 answer

    1. This product is not tested specifically for in vitro or in vivo use. A protocol is not currently available. However, a search of the literature reveals several publications that may be helpful. Please see the links below to review examples:

      A method to determine reactive oxygen species production in intestinal and liver cell cultures using the 2’,7’-dichlorodihydrofluorescein diacetate assay
      https://www.sciencedirect.com/science/article/pii/S2215016124000694

      Detection of Total Reactive Oxygen Species in Adherent Cells by 2’,7’-Dichlorodihydrofluorescein Diacetate Staining
      https://pmc.ncbi.nlm.nih.gov/articles/PMC7712457/

      Helpful?

  2. Is the product compatible with RFP transfected cells?

    1 answer

    1. Yes, RFP (Red Fluorescent Protein) and DCFH (2′,7′-Dichlorodihydrofluorescein) are spectrally compatible and can be used together for multicolor imaging or flow cytometry.

      Helpful?

  3. Hi, is the product suitable for ROS detection in a cell-free system? If not, could you please recommend a DCFH related product that works in a system without cells? Thanks

    1 answer

    1. This product as supplied is DCFH-DA and used for intracellular ROS detection. Please see the link below for understanding this process.
      https://pubs.acs.org/doi/10.1021/acs.analchem.7b00043

      For use in a cell-free system, deacetylation of DCFH-DA into DCFH must be performed before use.
      Please see the link below for this method.
      https://www.sciencedirect.com/science/article/pii/S0006295203000832?via%3Dihub

      Helpful?

  4. Is D6883 2´,7´-Dichlorofluorescin diacetate suitable for flow cytometry analyses?

    1 answer

    1. While the specifications for this probe do not include assaying for specific applications, there are many publications showing its use in flow cytometry applications:
      https://www.sciencedirect.com/science/article/pii/S0022030215000909
      https://iubmb.onlinelibrary.wiley.com/doi/pdf/10.1002/iub.1895
      https://www.tandfonline.com/doi/pdf/10.1080/21655979.2021.1994737
      https://journal.waocp.org/article_91422_6ccd2a19eb9159073edeb8f9ce0be660.pdf
      https://pmc.ncbi.nlm.nih.gov/articles/PMC7842666/

      Helpful?

  5. What is the protocol for measuring ROS using D6883? Specifically, what kind of media or buffer is used with this product?

    1 answer

    1. The specific protocol for measuring ROS using D6883 and the type of media or buffer to be used with this product are not available. However, typically, this product is added to a buffer solution shortly before assessing the fluorescent signal. For further guidance, refer to the following citations for relevant information:

      1. Novel Antioxidant Tripeptide "ACQ" Can Prevent UV-Induced Cell Death and Preserve the Number of Epidermal Stem Cells
      PMID: 26199677

      2. Detection of reactive oxygen species (ROS) by the oxidant-sensing probe 2',7'-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 7937
      PMID: 2057064

      These publications may provide insights into the protocol for using D6883 and the type of media or buffer used with this product.

      Helpful?

  6. What concentration of DCF can I use for A549 cells?

    1 answer

    1. This product has not been tested for use in cell culture. A search of the literature suggests working concentrations range from 5 µM to 0.1 mM. See the link below to a publication that specifically looked at A549 cells using several dyes, including DCFDA:
      https://pubs.rsc.org/en/content/articlehtml/2017/ra/c7ra01025g

      Helpful?

  7. What concentration of DCF can I use for THP-1 monocyte cell line?

    1 answer

    1. The optimal concentration of 2′,7′-Dichlorofluorescin diacetate (DCFDA) for use with THP-1 monocytes will need to be determined by the end-user. See the link below to a paper that reported using DCF at 40 uM with THP-1 cells.
      https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8961584/

      Helpful?

  8. Is it possible to view this probe under a fluorescence microscope if the probe is solubilised in DMSO? What are the excitation and absorption wavelengths?

    1 answer

    1. The excitation wavelength is 504 nm and the emission wavelength is 529 nm in ethanol. The use of DMSO as the solvent has not been determined, however, various publications suggest that it is suitable. See the link below for an example:
      https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7712457/

      Helpful?

  9. Protocol please! Thank you very much!

    1 answer

    1. It may be helpful to view the following articles as there is not a specific protocol for this product:

      https://www.sigmaaldrich.com/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/imaging-analysis-and-live-cell-imaging/mitochondrial-stress-and-ros

      https://www.sigmaaldrich.com/technical-documents/technical-article/research-and-disease-areas/cell-signaling/bioactive-small-molecules-for-nitric-oxide-cell-stress-research

      https://www.sigmaaldrich.com/search/d6883?focus=papers&page=1&perpage=30&sort=relevance&term=d6883&type=citation_search

      Helpful?

  10. Can the product be stored in powder form and if its possible what is the storage temp?

    1 answer

    1. This product should be stored in powdered format at -20°C.

      Helpful?

1–10 of 14 Questions  

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