Goat anti rabbit igg hrp antibody

Microneutralization assays were performed as previously with minor modifications⁷⁰ . Briefly, serially diluted sera and plasma were incubated with SARS-CoV-2 on Vero cells. Following incubation, cells were fixed, permeabilized, blocked and stained with rabbit anti-NP IgG antibody (Sino Biological, BJ, CN) (1:3000) followed by goat anti-rabbit IgG HRP antibody (Thermo Fisher Scientific) (1:3000). Plates were visualized with ABTS at 405 nm on a SpectraMAX 190. Readings were normalized with cell and virus controls and IC₅₀ readings were calculated using GraphPad Prism (La Jolla, CA, version 9.3.1) using the ‘log(inhibitor) vs normalized response’ function.

For Hematoxylin and Eosin (H&E) staining, fixed aorta tissues were successively dehydrated in 20 and 30% sucrose, embedded in O.C.T compound and then sectioned at 10 μm. The sections were stained with Hematoxylin (Solarbio, Beijing, China) and Eosin (Sangon, Shanghai, China) and observed under a microscope (40× and 200×, OLYMPUS, Tokyo, Japan). For immunohistochemistry (IHC) analysis, fixed samples were embedded in O.C.T compound, sectioned at 10 μm and fixed in pre-cooled acetone for 15 min. The sections were immersed in sodium citrate buffer for antigen retrieval and then incubated in 3% hydrogen peroxide for 15 min to eliminate endogenous peroxidase activity. Next, the sections were incubated in normal goat serum (Solarbio) for 15 min. Subsequently, the sections were incubated with α-SMA or calponin antibody (1:200 dilution, Proteintech) overnight at 4°C, followed by incubation with goat anti-rabbit IgG-HRP antibody (1:500 dilution, Thermo Fisher Scientific, Waltham, MA, U.S.A.) for 60 min at 37°C. The sections were stained with diaminobenzidine (DAB), counterstained with Hematoxylin and observed under a microscope (400×, OLYMPUS).