Immunohistochemical Detection of 8-OHdG

Dissected tissues were fixed in Bouin’s solution at 4°C overnight and processed in paraffin wax. Sections of 3 µm thickness were obtained on glass slides, deparaffinized, rehydrated and treated with decondensing buffer (25 mM DTT, 0.2% (v/v) Triton x-100 and 200 i.U./mL heparin in PBS) for 1 min at 37°C to increase antigen accessibility. Sections were further processed in the Lab Vision PT module (Thermo Fisher Scientific) and Autostainer 480S (Medac, Hamburg, Germany) as published previously [40 (link)]. The primary antibody against 8-OHdG (clone 15A3, Santa Cruz Biotechnology, Dallas, TX, USA) was used at a dilution of 1:200.