SARS-CoV-2 Nucleocapsid MAb (Clone 1035111) Summary
Accession # YP_009724397.2
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
SARS-CoV-2 Nucleocapsid in SARS-CoV-2 Infected Human Lung. SARS-CoV-2 Nucleocapsid was detected in immersion fixed paraffin-embedded sections of SARS-CoV-2 infected human lung (left, positive staining) and normal human lung (right, negative control) using Mouse Anti-SARS-CoV-2 Nucleocapsid Monoclonal Antibody (Catalog # MAB10474) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to SARS-CoV-2 infected cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of SARS-CoV-2 Nucleocapsid by Western Blot. Western blot shows recombinant SARS-CoV-2 Nucleocapsid protein. PVDF membrane was probed with 2 µg/mL of Mouse Anti-SARS-CoV-2 Nucleocapsid Monoclonal Antibody (Catalog # MAB10474) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for SARS-CoV-2 Nucleocapsid at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that are commonly comprised of four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) (1). While the S, E and M proteins build up the viral envelop, the N protein is involved transcription, replication and packaging of the viral RNA genome into a helical ribonucleocapsid (RNP) (2, 3). The SARS-CoV-2 N protein is a ~45 kDa protein composed of two independent structural domains connected by a linker region. The N-terminal region contains an RNA binding domain, the linker region interacts with the M protein and the C-terminal region contains a self-association domain (2,3). The SARS-CoV2 N protein shares 91% and 47% amino acid sequence identity with SARS-CoV-1 and MERS N protein, respectively. The SARS-CoV-2 N protein displays VSR (viral suppressor of RNA interference) activity in mammalian cells (4). In addition, the N protein is an abundant protein during coronavirus infection and displays high immunogenic activity (5, 6), so it has been used to develop serological diagnostic kit for Covid-19 IgM and IgG antibody tests (7).
- Wu, F. et al. (2020) Nature 579:265.
- Chang, C. K. et al. (2006) J. Biomed. Sci. 13:59.
- Hurst, K. R. et al. (2009) J. Virol. 83:7221.
- Mu, J. et al. (2020) Sci. China Life Sci. doi: 10.1007/s11427-020-1692-1.
- Che, X. Y. et al. (2004) J. Clin. Microbiol. 42:2629.
- Guan, M. et al. (2004) Clin. Diagn. Lab. Immunol. 11:287.
- Liu, W. et al. (2020) J. Clin. Microbiol. doi: 10.1128/JCM.00461-20.
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