Immunofluorescence assays were performed to examine protein expression in virus-infected Vero cells. Vero cells were infected at MOI 0.01 with PIV5, CVXGA1, CVXGA2, CVXGA3, CVXGA5, CVXGA13, or CVXGA14 for 3 days before being fixed with 80% methanol. The cells were incubated with mouse anti-PIV5 V/P monoclonal antibody (PK 366), rabbit anti-SARS-CoV-2 S (Sino Biological catalog no. 40150-R007), or SARS-CoV-2 N (ProSci catalog no. 35–579) antibodies at 1:500 in PBS + 3% bovine serum albumin (BSA) for 1 hr. Next, the cells were washed with PBS and incubated with goat α-mouse Cy3 (KPL) or goat α-rabbit Cy3 (KPL) at 1:500 in PBS + 3% BSA for 30 mins. The cells were washed with PBS and imaged with an EVOS M5000 microscope (Thermo Fisher Scientific).
Sars cov 2 n
Manufactured by ProSci
The SARS-CoV-2 N is a lab equipment product designed for the detection and identification of the SARS-CoV-2 nucleocapsid protein. It is a protein-based diagnostic tool used for research and analysis purposes.
Automatically generated - may contain errors
2 protocols using sars cov 2 n
1
Immunofluorescence Analysis of Virus-Infected Cells
2
SARS-CoV-2 nsp1 Protein Expression Analysis
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BHK-21 cells were transfected with the plasmids encoding Flag-GFP fusions of nsp1 proteins using Lipofectamine III (Invitrogen), according to the manufacturer’s protocol. At 18 h post transfection, the media were replaced with that supplemented with 10 μg/mL of Pur. Then, incubation continued for 15 min. Cells were harvested in PBS, pelleted via centrifugation, and lysed in standard protein loading buffer. The lysates were analyzed via Western blots using puromycin-specific MAbs (MABE343, Sigma) and infrared dye-labeled secondary Abs. Additional antibodies used: SARS-CoV-2 S2 (40590-T62, Sino Biological US Inc.), SARS-CoV-2 N (35-579, ProSci), SARS-CoV-2 Nsp1 (GTX135612, GeneTex), GFP (600-145-215, Rockland), β-actin (66009-1-Ig, Proteintech). The membranes were scanned on a LI-COR imager, and the data were processed, using the manufacturer’s software Empiria Studia. The data were normalized to the amount of β-actin in the samples.
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