Ab212057

Preparation for immunoelectron microscopy was performed as described previously (47 (link)). In brief, segments of liver and kidney were stained with rabbit anti–mouse BSG Ab (catalog ab212057; Abcam) or with PBS, followed by secondary hybridization with HRP-conjugated goat anti–rabbit IgG F(ab′)₂ (catalog 424141; Histofine; Nichirei Corporation). The stained tissues then were fixed with 1% glutaraldehyde, incubated with DAB for 30 minutes at room temperature, and washed twice with PBS. Tissues were postfixed in osmium tetroxide, dehydrated in alcohol, and embedded in epoxy resin (Quetol-812; Nissin EM Corporation). Ultrathin sections were examined using a JEM-1400 electron microscope (JOEL, Ltd.).

The tissue was fixed with 4% paraformaldehyde, embedded in paraffin and sectioned. The paraffin-embedded sections were dewaxed and rehydrated, and EDTA was used for antigen repair. At room temperature, sections were incubated with 3% H₂O₂ to quench endogenous peroxidase activity. Afterwards, samples were blocked with BSA and incubated overnight at 4℃ with specific primary antibodies, including anti-CD147 (ab212057, 1:200, Abcam, Cambridge, USA), anti-ACOX1 (ab184032, 1:200, Abcam), anti-FASN (ab128856, 1:200, Abcam). Later, samples were incubated for 1 h at 37°C with HRP-labeled goat anti-Rabbit secondary antibodies. The samples were then dyed with DAB and hematoxylin before sealing.