Key Product Details

Species Reactivity

SARS-CoV-2

Applications

Blockade of Receptor-ligand Interaction, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 1042445
View all Spike RBD Primary Antibodies »

Product Specifications

Immunogen

Chinese Hamster Ovary cell line CHO-derived  SARS-CoV Spike RBD
Arg306-Phe527
Accession # NP_0828851.1

Specificity

Detects SARS-CoV spike RBD in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

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

SARS-Cov-2 Spike RBD protein binding to ACE-2-transfected Human Cell Line is Blocked by SARS-Cov-2 Spike RBD Antibody

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 50 µg/mL of Mouse Anti-SARS-Cov-2 Spike RBD Monoclonal Antibody (Catalog # MAB108271) but not by (B) Mouse IgG1 Isotype Control (MAB002). Protein binding was detected with Mouse Anti-His APC-conjugated Monoclonal Antibody (IC050A). Staining was performed using our Staining Membrane-Associated Proteins protocol.

Detection of Spike RBD in CHO Chinese hamster ovary cell line transfected with SARS-CoV-2.

Spike RBD was detected in immersion fixed CHO Chinese hamster ovary cell line transfected with SARS-CoV-2 (positive staining) and CHO Chinese hamster ovary cell line (non-transfected, negative staining) using Mouse Anti-SARS-CoV-2 Spike RBD Monoclonal Antibody (Catalog # MAB108271) 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 cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for SARS-CoV-2 Spike RBD Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional flow cytometry test, 50 μg/mL of Mouse Anti-SARS-Cov-2 Spike RBD Antibody(Catalog # MAB108271) 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. 

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed CHO Chinese hamster ovary cell line transfected with SARS-CoV-2

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 was discovered in association with cases of severe acute respiratory syndrome (SARS) that infected more than 8,000 persons with over 900 fatalities worldwide in 2002-2003 (1). It belongs to a family of viruses known as coronaviruses that also include MERS and SARS-Cov2 that causes the global pandemic coronavirus disease 2019 (Covid-19). Coronavirus is commonly comprised of four structural proteins: Spike protein(S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) (1). SARS-CoV S Protein is a type-I trimerized membrane glycoprotein that mediates membrane fusion and viral entry. 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-CoV through interaction with a receptor binding domain (RBD) located at the C-terminus of S1 subunit (5, 6). Based on amino acid (aa) sequence homology, the RBD domain of SARS-Cov shares 73% and 24% homology with RBD domain of SARS-CoV2 and MERS, respectively. Before binding to the ACE-2 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 (7). Antibodies to S protein especially the RBD region of SARS-CoV have been shown to inhibit interaction with the ACE-2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (8).

References

  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. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
  8. Du, L. el al. (2009) Nat. Rev. Microbiol. 7:226.

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

NP_0828851.1

Additional Spike RBD Products

Product Documents for SARS-CoV-2 Spike RBD Antibody

Certificate of Analysis

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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

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