ECE2 Immunohistochemistry Staining Protocol

For IHC staining, sections were deparaffinized in xylene and rehydrated in alcohol, then endogenous peroxidase was inactivated by treatment with 3% H₂O₂ for 5 min. Then, antigen retrieval was performed in a pressure cooker with sodium citrate buffer (pH 6.0). After blocking with 5% goat serum for 30 min at room temperature, sections were incubated with rabbit ECE2 monoclonal antibody (1:200, Abcam, USA) overnight in a 4°C refrigerator. Sections were coupled with goat anti-mouse IgG-HRP secondary antibody (1:400, Abcam, USA) for 1 hour at room temperature, then each incubated section was stained with DAB reagent and finally counterstained with hematoxylin (12 (link)).