Western blot shows recombinant SARS-CoV-2 Spike S1 RBD. PVDF membrane was probed with 2 µg/mL of Mouse Anti-SARS-CoV-2 Spike RBD Monoclonal Antibody (Catalog # MAB10827) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Spike RBD at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of SARS-CoV-2 Spike S1 protein bound to ACE-2 in HEK293 Human Cell Line Transfected with Human ACE-2 and eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with human ACE-2 and eGFP was incubated with Recombinant SARS-CoV-2 Spike S1 Subunit His-Tag protein (10522-CV), then stained with (A) Mouse Anti-SARS-CoV-2 Spike S1 Monoclonal Antibody (Catalog # MAB10827) or (B) Mouse IgG2A Isotype Control Antibody (MAB003) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B). Staining was performed using our Staining Membrane-associated Proteins protocol.

Detection of Spike RBD in CHO Chinese hamster ovary cell line.

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 # MAB10827) 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

Flow Cytometry

0.25 µg/10⁶ cells
Sample: SARS-CoV-2 Spike S1 protein bound to ACE-2 in HEK293 Human Cell Line Transfected with Human ACE-2 and eGFP

Immunocytochemistry

3-25 µg/mL
Sample: immersion fixed CHO cell line transfected with SARS-CoV-2 Spike

Western Blot

2 µg/mL
Sample: Recombinant SARS-CoV-2 Spike S1 RBD

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.

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

Rota, P.A. et al. (2003) Science 300:1394.
Bosch, B.J. et al. (2003). J. Virol. 77:8801.
Belouzard, S. et al. (2009) Proc. Natl. Acad. Sci. USA 106:5871.
Millet, J.K. and G. R. Whittaker (2015) Virus Res. 202:120.
Li, W. et al. (2003) Nature 426:450.
Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
Ortega, J.T. et al. (2020) EXCLI J. 19:410.
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

UniProt

NP_0828851.1

Additional Spike RBD Products

Product Documents for SARS-CoV-2 Spike RBD Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

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

For research use only

Citations
Reviews

Calculators
Flow Cytometry

SARS-CoV-2 Spike RBD Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB10827

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

View all Spike RBD Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

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

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