Anti rat cd68 monoclonal antibody

To assess macrophage infiltration, the tissue sections were stained with anti-rat CD68 monoclonal antibody (1:100; AbD Serotec, Kidlington, UK) for 60 min at room temperature. To assess neutrophil infiltration, the tissue sections were stained with anti-rat myeloperoxidase (MPO) antibody (1:300, Thermo Fisher Scientific) overnight at 4°C. We photographed 10 random fields on a section from each rat and measured the ratio of stained areas in the mucosal layer using a digital image analyzer (WinROOF, Mitani Co., Fukui, Japan).

The tissue sections were stained with anti-rat type I collagen antibody (1:100,000; LSL, Tokyo, Japan) for 60 min at room temperature. To assess HSC activation, the tissue sections were stained with anti-rat α-SMA antibody (1:800, Thermo Fisher Scientific) for 30 min at room temperature. To assess the infiltration of Kupffer cells, the tissue sections were stained with anti-rat CD68 monoclonal antibody (1:50; AbD Serotec, Kidlington, United Kingdom) for 40 min at room temperature. We photographed 10 random fields on a section from each rat, and measured the stained areas from the entire liver cross-sectional area using a digital image analyzer (WinROOF).

To assess the activation of hepatic stellate cells (HSCs), tissue sections were stained with an anti-rat α-smooth muscle actin (SMA) antibody (1 : 800, Thermo Scientific, Waltham, MA, USA) for 30 minutes at room temperature. To assess the number of Kupffer cells (KCs), tissue sections were stained with an anti-rat CD68 monoclonal antibody (1 : 50; AbD Serotec, Kidlington, United Kingdom) for 40 minutes at room temperature. Ten random fields on a section from each rat were photographed, and stained areas were measured as percentage of the entire liver cross-sectional area.