Rabbit anti mers cov s protein polyclonal antibody

Indirect immunofluorescence assay (IFA) was used to detect S protein expression in rSRV9-MERS_S1-infected cells. NA cells were plated on coverslips in 35-mm-diameter dishes and infected with rSRV9-MERS_S1 or rSRV9 at a MOI = 1. At 24 h post-infection, cells were fixed in 4% paraformaldehyde (Solarbio Corporation, Beijing, China) at 4°C for 30 min and blocked with phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA) at RT for 1 h. Cells were incubated with rabbit anti-MERS-CoV S protein polyclonal antibody (Sino Biologicals, Beijing, China) and mouse anti-RABV G protein monoclonal antibody (Merck Millipore, Hong Kong, China) 1:100 diluted in PBS containing 1% BSA at 37°C for 1 h. Then the cells were washed 3 times with PBST and stained with 594-conjugated goat anti-rabbit antibody (Abcam, Shanghai, China, 1:500 diluted in PBS containing 1% BSA) and 488-conjugated goat anti-mouse antibody (Abcam, Shanghai, China, 1:1,000 diluted in PBS containing 1% BSA) at 37°C for 1 h. After being washed 3 times, cells were stained with an antifade mounting medium with DAPI (Vector Laboratories, Burlingame, CA, USA). Cells were analyzed with an Axio Scope.A1 microscope (Carl Zeiss MicroImaging GmbH, Göttingen, Germany), and representative images were obtained from the ZEN 2012 system (Carl Zeiss MicroImaging GmbH, Göttingen, Germany).

An indirect immunofluorescence assay (IFA) was used to detect S protein expression in MERS-CoV-infected cells. Huh7 cells infected with the virus (MOI = 0.0025, 0.005, or 0.01) were fixed using 4% paraformaldehyde (PFA) in PBS, and then the cells were permeabilized with 0.2% Triton X-100 in PBS. Cells were incubated with rabbit anti-MERS-CoV S protein polyclonal antibody (Sino Biologicals) at room temperature for 1 h and then stained with Fluor 594-conjugated goat anti-rabbit antibody (Abcam) (1:500 diluted in PBS containing 1% BSA); the nucleus was stained with DAPI. After being washed three times with PBS, the coverslips were mounted on glass slides. Cells were examined by confocal microscopy (Zeiss).