Lipid peroxidation was measured through the thiobarbituric acid reactive substances (TBARS) in plasma following a previously described protocol [18] (link). Reactive oxygen species (ROS) production was evaluated in the plasma through the fluorescence intensity of the redox-sensitive dye 2′,7′-dichlorodihydrofluorescein diacetate (DCFH, 100 μM, Sigma-Aldrich, St. Louis, MO, USA) (excitation and emission wavelengths of 480 and 535 nm, respectively) using SpectraMax M2e (Molecular Devices, St. Louis, MO, USA). The total antioxidant capacity (TAC) of plasma using 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) (Sigma-Aldrich) incubated with potassium persulphate (Sigma- Aldrich) to produce ABTS⋅+, a green/blue chromophore. The inhibition of ABTS⋅+ formation by antioxidants in the samples was expressed as Trolox equivalents, determined at 740 mm using a calibration curve plotted with different amounts of Trolox (Sigma-Aldrich). Nitrite levels were determined by Griess method as previously described [19] (link).
2 7 dichlorodihydrofluorescein diacetate dcfh
Manufactured by Merck Group
Sourced in United States
2',7'-dichlorodihydrofluorescein diacetate (DCFH) is a fluorogenic compound used in laboratory research. It serves as a precursor for the detection of reactive oxygen species.
Automatically generated - may contain errors
Lab products found in correlation
Spectramax m2e, by Molecular Devices (2 mentions)
Potassium persulphate, by Merck Group (1 mentions)
Trolox, by Merck Group (1 mentions)
Il 17a, by Thermo Fisher Scientific (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Il 10, by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
2 protocols using 2 7 dichlorodihydrofluorescein diacetate dcfh
1
Oxidative Stress Biomarkers Quantification
2
Evaluating Inflammatory Biomarkers and Oxidative Stress
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The concentrations of TNF-α, IL-6, IL-10, IL-17a and interferon-gamma (IFN-γ) (all from eBioscience, USA) in the supernatants of serum-stimulated cells and serum from participants were quantified by ELISA kits, according to the manufacturer’s protocols. Lipid peroxidation was measured through the TBARS in PBMCs following a previously described protocol69 (link) and expressed in MDA nmol/mL. Total thiol concentrations were measured in PBMCs using 5–5′-dithio-bis(2-nitrobenzoic acid) reagent9 . ROS production was evaluation in the cell lysate through the fluorescence intensity of the redox-sensitive dye 2′,7′-dichlorodihydrofluorescein diacetate (DCFH, 100 µM, Sigma-Aldrich) (excitation and emission wavelengths of 480 and 535 nm, respectively) using SpectraMax M2e (Molecular Devices, USA).
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