Polyclonal goat anti rabbit igg conjugated with horseradish peroxidase hrp

All the solvents were purchased from Sigma-Aldrich (Milan, Italy): ethanol (EtOH, gradient grade for HPLC, ≥99.9%), methanol (MeOH, gradient grade for HPLC, ≥99.9%), acetonitrile (ACN, LC-MS grade, ≥99.9%), water (H₂O, LC-MS grade, ≥99.9%), acetic acid (CH₃COOH, ≥98%), and formic acid (HCOOH, LC-MS grade, ≥98.5%). The oleuropein standard was purchased from Sigma-Aldrich (Milan, Italy) (98%), and the tomatine standard was purchased from Extrasynthese (Lyon, France) (≥98.5%). Cell culture media and antibiotics were purchased from Biosera (Nuaille, France). Phorbol 12-myristate 13-acetate (PMA) was obtained from Sigma-Aldrich (St. Louis, MO, USA). The rabbit polyclonal anti-malondialdehyde (MDA) antibody and MTT cell proliferation assay kit were purchased from Abcam [anti-malondialdehyde (MDA) antibody (ab27642)] (Cambridge, UK). Polyclonal goat anti-rabbit IgG conjugated with horseradish peroxidase (HRP) was obtained from Bio-Rad (Hercules, CA, USA). Protease and phosphatase inhibitors were obtained from Roche (Mannheim, Germany).

Fenton reagent preparation was conducted using H₂O₂ (Sigma-Aldrich Chemie GmbH, Mannheim, Germany) and ferrous sulfate (FeSO₄.7H₂O) (BDH Laboratory Supplies, Poole, UK). A variable concentration of H₂O₂ (2.5 mM, 5 mM, 10 mM) was used with a fixed concentration of iron sulphate (FeSO₄) (250 µM) to chemically generate HO^•. The procedure for topical application and its duration are specified in the figure legends, as applicable. Phosphatase and protease inhibitors were purchased from Roche (Mannheim, Germany). Rabbit polyclonal anti-MDA antibody was purchased from Abcam [anti-MDA antibody (ab27642)] (Cambridge, UK) and polyclonal goat anti-rabbit IgG conjugated with horseradish peroxidase (HRP) from Bio-Rad (Hercules, CA, USA). Rabbit polyclonal Dinitrophenyl-KLH antibody (anti-DNP) were procured from ThermoFisher scientific (Waltham, MA, USA).