Mers cov s rabbit polyclonal antibodies

Image cytometry methods were performed as previously described (Rosen et al., 2019 (link)). Briefly, BHK-21 cells were seeded into flat bottom black-walled Greiner 96-well plates and allowed to adhere overnight. On the following day, a DPP4 expression plasmid was transfected into the cells, using Lipofectamine 3000 reagent. Two days following DPP4 transfection, MERS-CoV S-2P was incubated with 4-fold serial dilutions of antibody (Fab or IgG) for 30 min at room temperature (RT). The mixture of MERS-CoV S and antibody was then added to the DPP4-expressing BHK-21 cells and incubated for 2 hours at RT. After incubation, cells were washed, fixed with 80% cold acetone, and rewashed. Subsequently, cells were stained with MERS-CoV S rabbit polyclonal antibodies (Sino Biological, Beijing, China) and then secondary goat anti-rabbit IgG H&L labeled with Alexa Fluor® 488 (AF488) was added. Finally, cell nuclei were stained with DAPI. Percent inhibition as a function of antibody concentrations was then plotted and analyzed via a one-site-fit Log IC₅₀ non-linear regression analysis. No inhibition (0%) was defined as MERS-CoV S binding to BHK-21 cells without the addition of antibody. Full inhibition (100%) was defined as MERS-CoV S binding to BHK-21 without DPP4 receptor.