H2dcfda dcf

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H2DCFDA (DCF) is a fluorogenic dye that measures the general level of oxidative stress in cells. It is a cell-permeant indicator that becomes fluorescent upon oxidation by reactive oxygen species.

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Lab products found in correlation

Fcs express 7, by De Novo Software (2 mentions) Facscanto 2, by BD (2 mentions) Accutase solution, by Merck Group (1 mentions)

Spelling variants of this product

H2DCFDA (1349 citations) CM-H2DCFDA (DCF, (6 citations) 2′,7′-dichlorofluorescein (DCF) diacetate (H2DCFDA) (2 citations)

3 protocols using h2dcfda dcf

Oxidative Stress Measurement using DCF

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Cells were treated for 15 min with H2DCFDA (DCF, Invitrogen) 1:5000 diluted in PBS. Data were acquired using a FACSCantoII (BD Bioscience) and were analyzed using FCS Express 7 (De Novo Software). Unstained samples were employed to set gates.

Mazzara P.G., Muggeo S., Luoni M., Massimino L., Zaghi M., Valverde P.T., Brusco S., Marzi M.J., Palma C., Colasante G., Iannielli A., Paulis M., Cordiglieri C., Giannelli S.G., Podini P., Gellera C., Taroni F., Nicassio F., Rasponi M, & Broccoli V. (2020). Frataxin gene editing rescues Friedreich’s ataxia pathology in dorsal root ganglia organoid-derived sensory neurons. Nature Communications, 11, 4178.

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Quantifying Microglia Oxidative Stress

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Microglia was treated for 15 m with H₂DCFDA (DCF, Invitrogen) 1:5000 diluted in PBS. Then cells were detached using Accutase Solution (Merck) 10 m at 37 °C. Data were acquired using a FACSCantoII (BD Bioscience) and were analyzed using FCS Express 7 (De Novo Software). Unstained samples were employed to set gates.

Bido S., Muggeo S., Massimino L., Marzi M.J., Giannelli S.G., Melacini E., Nannoni M., Gambarè D., Bellini E., Ordazzo G., Rossi G., Maffezzini C., Iannelli A., Luoni M., Bacigaluppi M., Gregori S., Nicassio F, & Broccoli V. (2021). Microglia-specific overexpression of α-synuclein leads to severe dopaminergic neurodegeneration by phagocytic exhaustion and oxidative toxicity. Nature Communications, 12, 6237.

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Measuring Sperm Intracellular ROS

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A FACSCalibur^® flow cytometer was used to analyze the levels of intracellular ROS of FT spermatozoa following the method described previously by [30 (link)]. In brief, FT samples were resuspended in 1 × DPBS. Afterward, 20 mM 2′, 7′-dichlorodihydrofluorescein diacetate (H₂DCFDA DCF, Invitrogen, Carlsbad, CA, USA) and PI were used to detect intracellular free radicals and the samples were incubated for 1 h at 25 °C in the dark. Afterward, the mean fluorescence intensity (MFI) of DCF was measured to evaluate intracellular mean H₂O₂ per viable sperm population.

Kim E., Almubarak A., Talha N., Yu I.J, & Jeon Y. (2021). The Use of κ-Carrageenan in Egg Yolk Free Extender Improves the Efficiency of Canine Semen Cryopreservation. Animals : an Open Access Journal from MDPI, 12(1), 88.

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