Cy3 conjugated donkey anti goat igg

For immunohistochemistry, deparaffinised sections were boiled in sodium citrate buffer (pH 6.0) for antigen retrieval. Endogenous peroxidase activity was inhibited by 3% hydrogen peroxide, and nonspecific binding was blocked using bovine serum albumin (BSA). Sections were then incubated overnight with the primary antibodies (Ki67: 1:750, #GB111141, Servicebio; CD31: 1: 600, #GB11063-2, Servicebio) at 4 °C. After incubation, sections were washed and incubated with the secondary antibody, HRP-conjugated goat anti-rabbit IgG (1:200, #G1213, Servicebio) at 23–25 °C for 50 min. Immunostaining was developed using a DAB solution (#G1211, Servicebio).
For immunofluorescence, deparaffinised sections were boiled for antigen retrieval and blocked for non-specific binding with BSA. The sections were then either incubated with fluorescent avidin conjugates (#A21370, Invitrogen) for 1 h at 23–25 °C for the identification of MCs, or overnight with the primary antibody at 4 °C (IL-33: 1:100, #AF3626, R&D Systems). After overnight incubation, sections were washed and incubated with the secondary antibody, Cy3-conjugated donkey anti-goat IgG (1:200, # GB21404, Servicebio), for 1 h at RT. Nuclei were stained with DAPI (#GDP1024, Servicebio). Fluorescence images were captured using a microscope (Nikon; Tokyo, Japan).