Dichlorodihydrofluorescein diacetate assay dcfh da

Dulbecco’s modified Eagle’s medium (DMEM), containing glucose at 4.5 mg/mL (25 mM), penicillin, streptomycin, trypsin–EDTA (ethylenediaminetetraacetic acid), FBS (Fetal Bovine Serum) and PBS (Phosphate-buffered saline) (without Ca and Mg) were provided by Gibco (San Diego, CA, USA), GSH/GSSG-Glo™ Assay kit, Caspase-Glo^® 3/7 Assay kit, Caspase-Glo^® 9 Assay kit were provided by Promega, Madison, WI, USA. MTT reagent was purchased from Sigma-Aldrich (St. Louis, MO, USA). Acridine orange and ethidium bromide were obtained from Sigma-Aldrich (St. Louis, MO, USA). Dichlorodihydrofluorescein diacetate assay (DCFH-DA) was provided by Sigma-Aldrich, St. Louis, MO, USA. SDS (Sodium dodecyl sulfate), TCA (Trichloroacetic acid), TBA (Thiobarbituric acid), Folin-Ciocalteu reagent were provided by Sigma-Aldrich and DTNB (Ellman’s Reagent) (5,5-dithio-bis-(2-nitrobenzoic acid)) by Serva. Cichoric acid, chlorogenic acid, caffeic acid, quercetin, DPPH (2,2-diphenyl-1-picrylhydrazyl), trolox, H₂O₂, horseradish peroxidase, phosphate buffer pH = 7 were purchased from Sigma-Aldrich Co. and used without purification. Methanol was purchased from Merck (Darmstadt, Germany).

All chemicals have an analytical purity and were used without further purification. Iron(III) chloride (FeCl₃·6H₂O), iron(III) sulfate (FeSO₄), trolox, H₂O₂, horse radish peroxide, phosphate buffer pH 7, Folin-Ciocalteu reagent, DPPH (2,2-diphenyl-1-picrylhydrazyl), 2,4,6-tripyridyl-s-triazine (TPTZ), trichloroacetic acid (TCA), thiobarbituric acid (TBA), sodium dodecyl sulphate (DSD) and dichlorodihydrofluorescein diacetate assay (DCFH-DA) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Ethanol and methanol were bought from Merck (Darmstadt, Germany). Dulbecco’s modified Eagle’s medium (DMEM) containing glucose at 4.5 mg/mL (25 mM), penicillin, streptomycin, trypsin–EDTA, fetal bovine serum (FBS) and phosphate buffered saline (PBS) without Ca and Mg were purchased from Gibco (San Diego, USA). DTNB (dithiobis-2-nitrobenzoic acid, Ellman's reagent) was provided by Serva Electrophoresis GmbH (Heidelberg, Germany). The fibroblasts cell line was obtained from American Type Culture Collection (ATCC).