Two hundred microliters of 20% trichloroacetic acid (TCA) were added to approximately 400 l of lysed semen samples in 0.8% SDS buffer, followed by centrifugation at 5.000 rpm for 10 min
Time points typically include 0, 5, 15, 30, 45, and 60 minutes, with additional points for slowly metabolized compounds
Cross-compartment protection : Hydrogen's ability to penetrate all cellular compartments complements the compartment-specific actions of water-soluble (vitamin C, B vitamins) and fat-soluble (vitamins A, E, D, K) antioxidants, ensuring no cellular region lacks protection
Briefly, HaCaT keratinocytes were seeded at a density of 10,000 cells per well in 200 l fresh culture medium (DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin solution) into 96-well flat-bottomed microtitre plates (Costar, Corning, USA) and incubated at 37 C in humidified 5% CO 2 using AutoFlow NU-5510 Direct Heat CO 2 Incubator (NuAire, USA) for 24 hours before treatment