For Western blotting, cells and tissues were lysed in RIPA buffer (Tris-HCl 25 mM, pH 7.6; NaCl 150 mM; NP-40 1%; sodium deoxycholate 1%; SDS 0.1%) with protease inhibitors and phosphatase inhibitors. Protein concentration was measured using bicinchoninic acid (BCA) (ComWin Biotech, China). Equal amounts of protein were separated by SDS-PAGE and transferred to a PVDF membrane which was incubated in 5% milk in Tris-buffered saline for 1 h at room temperature, followed by incubation with primary antibodies against ACE2 (Cell Signaling Technology, USA, 1:1000 dilution), FoxO1 (Cell Signaling Technology, USA, 1:1000 dilution), SARS-CoV-2 Nucleocapsid (NP) (SinoBiologicals, China, 1:1000 dilution), HA (Cell Signaling Technology, USA, 1:1000 dilution) and Flag (Cell Signaling Technology, USA, 1:1000 dilution) overnight at 4°C, and a secondary antibody (1:10000 dilution) conjugated with horseradish peroxidase (Cell Signaling Technology, USA) for 1 h at room temperature. Membranes were developed with chemiluminescent ECL reagents (Millipore, USA). The relative expression of target protein to the control β-actin were determined by densitometric analysis using ImageJ software (National Institutes of Health, USA).
Sars cov 2 nucleocapsid np
Manufactured by Sino Biological
Sourced in United States
The SARS-CoV-2 nucleocapsid (NP) is a recombinant protein produced by Sino Biological. It is a structural component of the SARS-CoV-2 virus and plays a crucial role in the viral genome packaging and assembly process.
Automatically generated - may contain errors
Lab products found in correlation
Ecl reagent, by Merck Group (1 mentions)
Flag tag (flag), by Cell Signaling Technology (1 mentions)
Horseradish peroxidase, by Cell Signaling Technology (1 mentions)
Foxo1, by Cell Signaling Technology (1 mentions)
Eclipse ti epifluorescence microscope, by Nikon (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Abcam (1 mentions)
3 protocols using sars cov 2 nucleocapsid np
1
Western Blotting of ACE2, FoxO1, and SARS-CoV-2 Proteins
2
Quantification of SARS-CoV-2 Nucleocapsid Protein
3
SARS-CoV-2 Neutralizing Antibody Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Δ6-RBD vaccines induced neutralizing activities against live SARS-CoV-2 WT or variants infection were detected using the plaque assay as described previously (25 (link)). The experiment was conducted in a BSL-3 laboratory at Shenzhen Center for Disease Control and Prevention. In brief, serum from each immunized mouse was diluted and mixed with the same volume of SARS-CoV-2 (100 PFU) and incubated at 37°C for 1 h. Thereafter, 200 μL of the virus-serum mixtures were transferred to pre-plated Vero E6 cells in 24-well plates. Inoculated cells were incubated at 37°C for two days. Then, Vero E6 cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. The cells were incubated sequentially with primary antibody against the SARS-CoV-2 nucleocapsid (NP) (SinoBiological) overnight at 4°C, horse radish peroxidase (HRP)-conjugated secondary antibody (Abcam), and TMB substrate (KPL). The plaque reduction neutralizing antibody titer (PRNT50) was defined as the minimal serum dilution that suppressed > 50% of viral plaques.
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