Nucleocapsid Antibody (1051436) [Unconjugated]

Western blot shows the detection of recombinant HCoV-OC43-N. PVDF membrane was probed with 1 µg/mL of Mouse Anti-HCoV-OC43 Nucleocapsid Monoclonal Antibody (Catalog # MAB11289) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Nucleocapsid at approximately 51 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Nucleocapsid was detected in immersion fixed Transfected & Wild Type HEK293 Human Embryonic Kidney Cell Line using Mouse Anti-HCoV-OC43 Nucleocapsid Monoclonal Antibody (Catalog # MAB11289) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surface and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

HEK293 cells transfected with HCOV-OC43-N or HEK293 cells transfected with an irrelevant protein were stained with Mouse Anti-HCoV-OC43 Nucleocapsid Monoclonal Antibody (Catalog # MAB11289, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

• Reactivity: V
• Applications: WB, Flow, ICC/IF
• Clone: 1051436
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Nucleocapsid Antibody (1051436) [Unconjugated] Summary

• Immunogen: Sf-21 (baculovirus)-derived hcov-oc43 Nucleocapsid
  Met1-Ile448
  Accession # YP_009555245.1
• Specificity: Detects HCoV-OC43 Nucleocapsid in direct ELISA.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 0.5 ug/10^6 cells
  • Immunocytochemistry 8-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Nucleocapsid Antibody (1051436) [Unconjugated]

• N Protein
• Nucleocapsid protein
• Nucleocapsid
• ORF9a protein

Background

HCoV-OC43, a virus first isolated in 1960's that accounts for ~ 20% of the common cold, belongs to a family of viruses known as coronaviruses that are commonly comprised of a large plus-strand RNA genome and four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) (1, 2). Other well-known human coronaviruses include three viruses that cause relatively mild respiratory disease: HCoV-229E, HCoV-HKU1 and HCov-NL63, plus three viruses that cause the Severe Acute Respiratory Syndrome (SARS-CoV), the Middle East Respirator Syndrome (MERS-CoV), and the global pandemic Covid-19 (SARS-CoV2). 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) (3, 4). The CoV-OC43 N protein is a ~50 kDa protein composed of two independent structural domains connected by a linker region. Both the N-terminal and the linker regions contain RNA binding domains, while the C-terminal region is responsible for the oligomerization of the N protein (5). The CoV‑OC43 N protein shares 64% amino acid sequence identity with CoV-HKU1 N protein. the N protein is an abundant protein during coronavirus infection and displays high immunogenic activity. Cross activity of antibodies among different strains should be rigorously tested when designing serological diagnostic kits (6, 7).

1. St-Jean, J.R. et al. (2004) J. Virol. 78:8824.
2. Vabret, A. et al. (2003) Fr Clin Infect Dis. 36:985.
3. Chang, C.K. et al. (2006) J. Biomed. Sci. 13:59.
4. Hurst, K.R. et al. (2009) J. Virol. 83:7221.
5. Huang, C.Y. et al. (2009) Protein Sci. 18:2209.
6. Chan, K.H. et al. (2005) Clin Diagn Lab Immunol. 12:1317.
7. Mourez, T. et al. (2007) J. Virol Methods. 139:175.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • YP_009555245.1()