Peroxidase dab 3 3 diaminobenzidine

Immunohistochemistry was performed using an avidin–biotin complex immunoperoxidase technique. Human-specific antibodies with no cross-reactivity to mouse antigens were used to label the following markers: α smooth muscle actin (α-SMA, Cat. No. NB600-536, Novusbio, USA); albumin (Cat. No. NBP2-21669, Novusbio, USA); and Von Willebrand factor (Cat. No. NB600-586, Novusbio, USA). The anti-OxPhos complex IV subunit I monoclonal antibody (Cat. No. 459600, Invitrogen, USA) and the rabbit monoclonal Anti-f4/80 mouse antibody (Cat. No. AB111101, Abcam) were also used. We used a preformed streptavidin–biotin–peroxidase complex and peroxidase-DAB (3,3′-diaminobenzidine) (Dako, Denmark) according to the manufacturer’s instructions. Sections were counterstained with Mayer’s hematoxylin and mounted using DPX medium. Positive and negative control slides for each marker were included in each run. As a negative control, a liver tissue section was processed as described but with the primary antibody omitted.

Immunohistochemistry (IHC) was performed on 5 µ-thick sections of formalin-fixed and paraffin-embedded tissue samples. Sections on charged slides are treated with Rabbit polyclonal anti-CD117 (c-kit) antibodies (CD 117, Cat. No. A4502, Dako, Denmark) as the primary antibody to label the c-kit positive cells and evaluate their density and configuration. Goat anti-mouse biotinylated immunoglobulins/HRP (Cat. No. P0447, Dako, Denmark) were utilized as the secondary antibody. Streptavidin–biotin–peroxidase complex and peroxidase-DAB (3,3′-diaminobenzidine) (Dako, Denmark) were applied. All were performed according to the manufacturer’s instructions. Positive and negative control slides were included in each run.