SARS-CoV-2 Spike RBD Antibody (1056608)

R&D Systems | Catalog # MAB11227

R&D Systems

Key Product Details

Species Reactivity

SARS-CoV-2

Applications

Blockade of Receptor-ligand Interaction, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 1056608
View all Spike RBD Primary Antibodies »

Product Specifications

Immunogen

Recombinant SARS-CoV-2 B.1.1.529 (Omicron) Spike RBD domain.
Arg319-Phe541
Accession # YP_009724390.1

Specificity

It detects SARS-CoV-2 Omicron BA.1 in Direct ELISA. In ELISA, this antibody does not detect SARS-CoV-2 Alpha, Gamma or Delta variants.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for SARS-CoV-2 Spike RBD Antibody (1056608)

Detection of Spike RBD in HEK293 (positive) and HEK293 wild type (negative) cells.

Spike RBD was detected in immersion fixed HEK293 (positive) and HEK293 wild type (negative) cells using Mouse Anti-SARS-CoV-2 Spike RBD Monoclonal Antibody (Catalog # MAB11227) at 3 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 cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Blocking of Spike RBD in HEK293 cells transfected with Human ACE-2 and eGFP cells by Flow Cytometry.

In a functional flow cytometry test, Recombinant SARS-Cov-2 Spike RBD His-tagged protein (10500-CV) binds to HEK293 human embryonic kidney cell line transfected with recombinant human ACE-2 and eGFP. (A) Binding is blocked by 25µg/mL of Mouse Anti-SARS-CoV-2 Spike RBD Monoclonal Antibody (Catalog # MAB11227) but not by (B) Mouse IgG1 Isotype Control (Catalog # MAB002). Protein binding was detected with His Tag Allophycocyanin Mab (Catalog # IC050A). View our protocol for Staining Membrane-associated Proteins.

Applications for SARS-CoV-2 Spike RBD Antibody (1056608)

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional flow cytometry test, 25 μg/mL of Mouse Anti-SARS-CoV-2 Spike RBD Monoclonal Antibody (Catalog # MAB11227) will block the binding of Recombinant SARS-Cov-2 Spike RBD His-tagged protein (Catalog # 10500-CV) to HEK293 human embryonic kidney cell line transfected with recombinant human ACE-2 and eGFP

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed HEK293 (positive) and HEK293 wild type (negative) cells

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Spike RBD

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‑CoV 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). The SARS-CoV-2 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). In SARS-CoV-2, as with most coronaviruses, proteolytic cleavage of the S protein into 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 (3-5). A receptor binding domain (RBD) in the C-terminus of the S1 subunit has been identified and the RBD of SARS-CoV-2 shares 73% amino acid (aa) identity with the RBD of the SARS-CoV-1, but only 22% aa identity with the RBD of MERS‑CoV (6, 7). The low aa sequence homology is consistent with the finding that SARS and MERS‑CoV bind different cellular receptors (8). The RBD of SARS-CoV-2 binds a metallopeptidase, angiotensin-converting enzyme 2 (ACE-2), similar to SARS-CoV-1, but with much higher affinity and faster binding kinetics (9). Before binding to the ACE-2 receptor, structural analysis of the S1 trimer shows that only one of the three RBD domains is in the "up" conformation. This is an unstable and transient state that passes between trimeric subunits but is nevertheless an exposed state to be targeted for neutralizing antibody therapy (10). Polyclonal antibodies to the RBD of the SARS-CoV-2 protein have been shown to inhibit interaction with the ACE-2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (11). There is also promising work showing that the RBD may be used to detect presence of neutralizing antibodies present in a patient's bloodstream, consistent with developed immunity after exposure to the SARS-CoV-2 (12). Several emerging SARS-CoV-2 genomes have been identified including the Omicron, or B.1.1.529, variant. First identified in November 2021 in South Africa, the Omicron variant quickly became the predominant SARS-CoV-2 variant and is considered a variant of concern (VOC). The Omicron variant contains 15 mutations in RBD domain that potentially affect viral fitness and transmissibility. The majority of the mutations are involved in ACE-2 binding and Omicron binds ACE-2 with greater affinity, potentially explaining its increased transmissibility (13, 14). Several of these mutations are also identified in facilitating immune escape and reducing neutralization activity to several monoclonal antibodies (13). Additionally, a series of new mutations are present in the RBD which have unknown impacts on receptor binding or antibody neutralization.

References

  1. Wu, F. et al. (2020) Nature 579:265.
  2. Tortorici, M.A. and D. Veesler (2019) Adv. Virus Res. 105:93.
  3. Bosch, B.J. et al. (2003) J. Virol. 77:8801.
  4. Belouzard, S. et al. (2009) Proc. Natl. Acad. Sci. 106:5871.
  5. Millet, J.K. and G.R. Whittaker (2015) Virus Res. 202:120.
  6. Li, W. et al. (2003) Nature 426:450.
  7. Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
  8. Jiang, S. et al. (2020) Trends. Immunol. https://doi.org/10.1016/j.it.2020.03.007.
  9. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
  10. Wrapp, D. et al. (2020) Science 367:1260.
  11. Tai, W. et al. (2020) Cell. Mol. Immunol. 17:613.
  12. Okba, N.M.A. et al. (2020). Emerg. Infect. Dis. https://doi.org/10.3201/eid2607.200841.
  13. Shah, M. and H.G. Woo (2022) Front. Immunol. https://doi.org/10.3389/fimmu.2021.830527.
  14. Lupala, C.S. et al. (2021) Biochem. Biophys. Res. Comm. (2022) https://doi.org/10.1101/2021.12.10.472102.

Long Name

Spike Receptor Binding Domain

Entrez Gene IDs

3200426 (HCoV-HKU1); 14254594 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)

Gene Symbol

S

UniProt

YP_009724390.1

Additional Spike RBD Products

Product Documents for SARS-CoV-2 Spike RBD Antibody (1056608)

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Product Specific Notices for SARS-CoV-2 Spike RBD Antibody (1056608)

For research use only

Related Research Areas

Infectious Diseases

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