Peroxidase conjugated goat anti mouse igg secondary antibody

Immunocytochemical staining was performed, in order to determine the purity of the stromal cells following isolation. The cells were cultured on cover slides in 30 mm culture dishes (Corning China, Shanghai, China), and fixed with 4% paraformaldehyde. Cells were permeabilized with 0.5% Triton X-100 (Shanghai Solarbio Bioscience & Technology Co., Ltd., Shanghai, China) for 10 min, rinsed with PBS three times, incubated with 3% H₂O₂ for 15 min to remove endogenous peroxidase and incubated for 30 min with 5% BSA for antigen blocking (Beyotime Institute of Biotechnology). Primary antibodies targeting mouse monoclonal cytokeratin 18 (cat. no. sc-32329; 1:400 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and mouse monoclonal vimentin (cat. no. sc-373717; 1:400 dilution; Santa Cruz Biotechnology, Inc.) were incubated with cells for 2 h at room temperature followed by incubation with peroxidase-conjugated goat anti-mouse IgG secondary antibody (cat. no. ZDR-5307; ZSGB-BIO, Beijing, China) for 40 min at room temperature, as previously described (19 (link)). Images were captured using a Leica DM3000 microscope (Leica Microsystems GmbH, Wetzlar, Germany).