SARS-CoV-2 Nucleocapsid Antibody

Catalog #: MAB104741 Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
MAB104741-100
MAB104741-SP
Detection of SARS-CoV-2 Nucleocapsid by Western Blot.
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SARS-CoV-2 Nucleocapsid Antibody Summary

Specificity
Detects SARS-CoV-2 Nucleocapsid in ELISAs and Western blots. No cross-reactivity with MERS Nucleocapsid is observed in Western blots.
Source
Monoclonal Mouse IgG1 Clone # 1035145
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Spodoptera frugiperda, Sf 21-derived SARS-CoV-2 Nucleocapsid protein
Met1-Ala419
Accession # YP_009724397.2
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
Recombinant SARS-CoV-2 Nucleocapsid protein
ELISA

This antibody functions as an ELISA capture antibody when paired with Mouse Anti-SARS-CoV-2 Nucleocapsid Monoclonal Antibody (Catalog # MAB104742).

This product is intended for assay development on various assay platforms requiring antibody pairs.

 
Immunohistochemistry
5-25 µg/mL
Immersion fixed paraffin-embedded sections of SARS-CoV-2 infected human lung

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.

Scientific Data

Western Blot Detection of SARS-CoV-2 Nucleocapsid antibody by Western Blot. View Larger

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 # MAB104741) 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.

SARS-CoV-2 Nucleocapsid ELISA Standard Curve. Recombinant SARS-CoV-2 Nucleocapsid protein was serially diluted 2-fold and captured by Mouse Anti-SARS-CoV-2 Nucleocapsid Monoclonal Antibody (Catalog # MAB104741) coated on a Clear Polystyrene Microplate (DY990). Mouse Anti-SARS-CoV-2 Nucleocapsid Monoclonal Antibody (MAB104742) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998) followed by Substrate Solution (DY999) and stopping the enzymatic reaction with Stop Solution (DY994).

Immunohistochemistry View Larger

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 # MAB104741) 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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.

Background: Nucleocapsid

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).

References
  1. Wu, F. et al. (2020) Nature 579:265.
  2. Chang, C. K. et al. (2006) J. Biomed. Sci. 13:59.
  3. Hurst, K. R. et al. (2009) J. Virol. 83:7221.
  4. Mu, J. et al. (2020) Sci. China Life Sci. doi: 10.1007/s11427-020-1692-1.
  5. Che, X. Y. et al. (2004) J. Clin. Microbiol. 42:2629.
  6. Guan, M. et al. (2004) Clin. Diagn. Lab. Immunol. 11:287.
  7. Liu, W. et al. (2020) J. Clin. Microbiol. doi: 10.1128/JCM.00461-20.
Long Name
Nucleocapsid Protein
Entrez Gene IDs
918763 (HCoV-229E); 3200423 (HCoV-HKU1); 2943504 (HCoV-NL63); 39105221 (HCoV-OC43); 14254601 (MERS-CoV); 1489678 (SARS-CoV); 43740575 (SARS-CoV-2)
Alternate Names
N Protein; Nucleocapsid

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