Rabbit monoclonal anti ki67 antibody

For histological analysis, liver tissue was cut into 5-mm sections, deparaffinized with xylene, and hydrated through graded ethanol. Ki67 was stained using Rabbit monoclonal anti-Ki67 antibody (Thermo Scientific) and diluted 1:100; F4/80 was stained using rat monoclonal anti-F4/80 antibody (MCA497; Serotec, Raleigh, NC) diluted 1:200. Neutrophils were stained using Rat anti- Mouse Gr-1 antibody (MCA2387, Serotec) diluted 1:200; γ-H2AX was stained using mouse antibody to phospho-H2AX diluted 1:100 (05-636; Upstate). Rat anti Mouse CD3 antibody diluted 1:300 (MCA500G, Serotec). Rabbit anti-phospho-Histone H3 (Ser10) antibody diluted 1:600 (06-570, Upstate). α -Smooth muscle antibody diluted 1:300 (A2547, Sigma Aldrich). For all staining, we used a conjugated horseradish peroxidase secondary Ab (anti-mouse and -rabbit [Envision; Dako] and anti-rat [Histifine; Nichirei, Osaka, Japan]) for 1 hour and developed it with AEC for 15 minutes. TUNEL staining was performed with an in situ Cell Death Detection Kit (Roche Diagnostics). H&E and Masson trichrome staining were performed according to accepted protocols. For the quantitative assessment of F4/80 staining, we used the Ariol system (Genetix USA Inc., San Jose, CA) for automated cell image capture and analysis.

One out of 20 sections was taken starting from the beginning of the lateral ventricle up to the end of the hippocampal formation (n = 16–20 sections). Sections were incubated overnight at 4 °C with rabbit monoclonal anti-Ki67 antibody (1:100; Thermo Scientific). Detection was performed with a HRP-conjugated anti-rabbit secondary antibody (1:200; Jackson Immunoresearch) and DAB kit (Vector Laboratories).