Ferrous sulfate feso4.7h2o

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).

Fenton’s reagent was prepared using hydrogen peroxide (H₂O₂) (Sigma-Aldrich Chemie GmbH, Germany) and ferrous sulfate (FeSO₄.7H₂O) (BDH Laboratory Supplies, United Kingdom). A fixed concentration of FeSO₄ (500 μM) and a variable concentration of H₂O₂ (100 μM/1 mM) was used_. Spin trapping reagent, POBN [α-(4-Pyridyl 1-oxide)-N-tert-butylnitrone] was purchased (Sigma-Aldrich Chemie GmbH, Germany).