Key Product Details

Species Reactivity

Validated:

SARS-CoV-2

Cited:

Human, Plant - Nicotiana benthamiana, Primate - Cercopithecus aethiops (African Green Monkey)

Applications

Validated:

Western Blot

Cited:

Western Blot, Bioassay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 1034522
View all Spike RBD Primary Antibodies »

Product Specifications

Immunogen

Recombinant SARS-CoV-2 Spike Protein, S1 Subunit, Receptor Binding Domain (RBD)
Arg319-Phe541
Accession # YP_009724390

Specificity

Detects SARS-CoV-2 Spike Protein S1 Receptor Binding Domain (RBD) in direct ELISAs and Western blots. Detects SARS-CoV-2 B.1.1.529 S RBD (Omicron Variant) in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for SARS-CoV-2 Spike RBD Antibody

Detection of SARS-CoV-2 Spike RBD by Western Blot.

Western blot shows recombinant SARS-CoV-2 Spike RBD protein. PVDF membrane was probed with 2 µg/mL of Mouse Anti-SARS-CoV-2 Spike RBD Monoclonal Antibody (Catalog # MAB10540) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for SARS-CoV-2 Spike RBD at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of SARS-CoV-2 Spike RBD by Western Blot

Detection of SARS-CoV-2 Spike RBD by Western Blot

A conjugate vaccine targeting the RBD domain of SARS-CoV-2(A–C) Schematic representation of the RBD (A), CDP (B), and produced CDP-RBD (28.9 kDa) and RBD (22.8 kDa) vaccine proteins (C).(B) CDP is composed of several highly immunogenic clusters originating from three different bacterial proteins (cell division protein ZapB, type-1 fimbrial protein TFP, and small heat shock protein IbpA).(C) The theoretical isoelectric point (pI) and the predicted solubility score (Sol) for each vaccine protein.(D) Coomassie staining after SDS-PAGE (left) and anti-RBD Western blot (right) analysis showing RBD and CDP-RBD.(E) Size-exclusion chromatogram (10/300 GL Superdex 75, Cytiva) showing purified CDP-RBD (black arrow). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35813877), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of SARS-CoV-2 Spike RBD by Western Blot

Detection of SARS-CoV-2 Spike RBD by Western Blot

A conjugate vaccine targeting the RBD domain of SARS-CoV-2(A–C) Schematic representation of the RBD (A), CDP (B), and produced CDP-RBD (28.9 kDa) and RBD (22.8 kDa) vaccine proteins (C).(B) CDP is composed of several highly immunogenic clusters originating from three different bacterial proteins (cell division protein ZapB, type-1 fimbrial protein TFP, and small heat shock protein IbpA).(C) The theoretical isoelectric point (pI) and the predicted solubility score (Sol) for each vaccine protein.(D) Coomassie staining after SDS-PAGE (left) and anti-RBD Western blot (right) analysis showing RBD and CDP-RBD.(E) Size-exclusion chromatogram (10/300 GL Superdex 75, Cytiva) showing purified CDP-RBD (black arrow). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35813877), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for SARS-CoV-2 Spike RBD Antibody

Application
Recommended Usage

Western Blot

2 µg/mL
Sample: Recombinant SARS-CoV-2 Spike RBD protein

Reviewed Applications

Read 1 review rated 4 using MAB10540 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma 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 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). In SARS-CoV-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 (3-5). Based on structural biology studies, the receptor binding domain (RBD), located in the C-terminal region of S1, can be oriented either in the up/standing or down/lying state (6). The standing state is associated with higher pathogenicity and both SARS-CoV-1 and MERS can access this state due to the flexibility in their respective RBDs. A similar two-state structure and flexibility is found in the SARS-CoV-2 RBD (7). Based on amino acid (aa) sequence homology, the SARS-CoV-2 S1 subunit RBD has 73% identity with the RBD of the SARS-CoV-1 S1 RBD, but only 22% homology with the MERS S1 RBD. The low aa sequence homology is consistent with the finding that SARS and MERS bind different cellular receptors (8). The S Protein of the SARS-CoV-2 virus, like the SARS-CoV-1 counterpart, binds Angiotensin-Converting Enzyme 2 (ACE2), but with much higher affinity and faster binding kinetics (9). Before binding to the ACE2 receptor, structural analysis of the S1 trimer shows that only one of the three RBD domains in the trimeric structure 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 ACE2 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 virus (12). Lastly, it has been demonstrated the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (13, 14).

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. Yuan, Y. et al. (2017) Nat. Commun. 8:15092.
  7. Walls, A.C. et al. (2010) Cell 180:281.
  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. https://doi.org/10.1016/j.it.2020.03.007.
  12. Okba, N. M. A. et al. (2020). Emerg. Infect. Dis. https://doi.org/10.3201/eid2607.200841.
  13. Wang, X. et al. (2020) https://doi.org/10.1038/s41423-020-0424-9.
  14. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1.

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

Additional Spike RBD Products

Product Documents for SARS-CoV-2 Spike RBD Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for SARS-CoV-2 Spike RBD Antibody

For research use only

Related Research Areas

Infectious Diseases

Citations for SARS-CoV-2 Spike RBD Antibody

Customer Reviews for SARS-CoV-2 Spike RBD Antibody (1)

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  • SARS-CoV-2 Spike RBD Antibody
    Name: David Krones
    Application: Western Blot
    Sample Tested: Recombinant protein
    Species: Hamster
    Verified Customer | Posted 12/14/2021
    Antibody was diluted in Tris buffered buffer with 0,5% Tween 20 added and incubated overnight at 4°C under constant shaking. Antibody was diluted as instructed and final dilution was 1:1000 equaling 0,5 µg/ml concentration.
    Samples were boiled for 10 min in 4x Laemmli buffer and subjected to 10% SDS-PAGE.
    SARS-CoV-2 Spike RBD Antibody MAB10540

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