SARS-CoV-2 Spike S2 Antibody (2812C) [Unconjugated]

Western blot shows recombinant SARS-CoV-2 Spike S2 protein. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-SARS-CoV-2 Spike S2 Subunit Monoclonal Antibody (Catalog # MAB10850) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Spike S2 Subunit at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Spike S2 Subunit was detected in immersion fixed HEK293 human embryonic kidney cell line transfected with SARS-CoV-2 (positive staining) and HEK293 human embryonic kidney cell line (non-transfected, negative staining) using Rabbit Anti-SARS-CoV-2 Spike S2 Subunit Monoclonal Antibody (Catalog # MAB10850) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Spike S2 Subunit was detected in immersion fixed paraffin-embedded sections of SARS-CoV-2 infected human lung using Rabbit Anti-SARS-CoV-2 Spike S2 Subunit Monoclonal Antibody (Catalog # MAB10850) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). 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 immunoreactive profiles scattered throughout the tissue. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western lane view shows recombinant SARS-CoV-2 (positive sample) and OC43 coronavirus lysate (negative control), loaded at 0.2 mg/mL. A specific band was detected for the Spike Protein at approximately 268 kDa (as indicated) using 20 µg/mL of Rabbit Anti-SARS-CoV-2 Spike S2 Subunit Monoclonal Antibody (Catalog # MAB10850) . This experiment was conducted under non-reducing conditions and using the 12-230 kDa separation system.

• Reactivity: V
• Applications: WB, Simple Western, IHC, ICC/IF
• Clone: 2812C
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

SARS-CoV-2 Spike S2 Antibody (2812C) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: E. coli-derived recombinant human SARS-CoV-2 Spike S2 Subunit
  Met697-Pro1213
• Specificity: Detects SARS-CoV-2 Spike S2 Subunit in direct ELISAs.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-25 ug/mL
  • Immunohistochemistry 3-25 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 0.5 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 SARS-CoV-2 Spike S2 Antibody (2812C) [Unconjugated]

• COVID-19
• S2 protein
• SARS-CoV-2 Spike
• SARS-CoV-2
• Spike glycoprotein Subunit 2
• Spike protein S2
• spike protein
• Spike S2 Subunit
• structural protein
• surface glycoprotein

Background

SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that also include MERS and SARS-CoV-1. Coronaviruses are commonly comprised of four structural proteins: Spike protein(S), Envelope protein (E), Membrane protein (M) and Nucleocapsid protein (N) (1). SARS-CoV-2 Spike Protein (S Protein) is a glycoprotein that mediates membrane fusion and viral entry. The S protein is homotrimeric, with each ~180-kDa monomer consisting of two subunits, S1 and S2 (2). As with most coronaviruses, proteolytic cleavage of the S protein into two distinct peptides, S1 and S2 subunits, is required for activation. The S1 subunit is focused on attachment of the protein to the host receptor while the S2 subunit is involved with cell fusion (2-4). A metallopeptidase, angiotensin-converting enzyme 2 (ACE-2), has been identified as a functional receptor for SARS-CoV2, similar to SARS-CoV-1, through interaction with a receptor binding domain (RBD) located at the C-terminus of S1 subunit (5, 6). The S2 subunit of SARS-CoV-2 shares 90% and 41% amino acid sequence identity with the S2 subunit of SARS-CoV-1 and MERS, respectively. It has been demonstrated the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (7, 8).

1. Rota, P.A. et al. (2003) Science 300:1394.
2. Bosch, B.J. et al. (2003). J. Virol. 77:8801.
3. Belouzard, S. et al. (2009) Proc. Natl. Acad. Sci. USA 106:5871.
4. Millet, J.K. and G. R. Whittaker (2015) Virus Res. 202:120.
5. Li, W. et al. (2003) Nature 426:450.
6. Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
7. Wang, X. et al. (2020) https://doi.org/10.1038/s41423-020-0424-9.
8. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v.

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.