Mouse anti gr 1 antibody

To visualize neutrophil infiltration in the liver, liver tissue sections were stained for Gr-1. Paraffin-embedded sections were dewaxed in xylene and rehydrated in a graded series of ethanol. Slides were incubated in 0.92% citric acid buffer (Vector Laboratories, Burlingame, CA) at 95°C for 15 min. Slides were then cooled to room temperature and incubated with 2% H₂O₂ in 60% methanol and blocked in 5% normal rat serum/Tris buffered saline (TBS), after which they were incubated with mouse anti-GR-1 antibody (1:50 dilution) (Cat 108402, Biolegend, San Diego CA) in 2% normal horse serum/TBS with 0.02% Triton X-100 at 4°C overnight. The detection was carried out as per the instructions provided by a commercially available immunohistochemistry kit with NovaRED substrate (Vector Laboratories). Gr-1 positive cells were counted using ImageJ software. For each section, 5 representative images were taken throughout and the number of Gr-1 positive cells was averaged across the section.