Goat anti human fix antibody

For immunostaining of human FIX, murine liver samples were harvested after 9 weeks of hepatic gene transfer. Samples were embedded in OCT media (Polyfreeze; Sigma–Aldrich), sectioned at 10 μm thickness, and fixed in 4% paraformaldehyde for 15 min at room temperature. Slides were washed with PBS and blocked in a solution containing 10% normal donkey serum (Santa Cruz Biotechnology), 0.2% Triton X-100 (Sigma–Aldrich) diluted in PBS for 1 h at room temperature. Subsequently, sections were incubated with goat anti-human FIX antibody (1:100; Affinity Biologicals, Hamilton, ON, Canada) overnight at 4°C. After washing thrice, the slides were incubated with donkey anti-goat Cy3 antibody (Jackson ImmunoResearch, West Grove, PA) at dilution of 1:500 for 1.5 h at room temperature. Sections were washed thrice and mounted with Fluoroshield™ with DAPI (Sigma–Aldrich). Images were acquired by Leica DMi8 confocal microscope (Wetzlar, Germany).