Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein, CF

Sf21 Insect Cell Expressed
Catalog # Availability Size / Price Qty
10522-CV-100
10522-CV-01M
Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein Bioactivity
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Product Details
Citations (4)
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Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.30 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag ( (Catalog # 933-ZN).
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived sars-cov-2 Spike S1 Subunit protein
Val16-Pro681, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Val16, sequence confirmed by mass spectrometry
Predicted Molecular Mass
75 kDa
SDS-PAGE
78-92 kDa, under reducing conditions

Product Datasheets

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10522-CV

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

10522-CV

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 200 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Bioactivity Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein Bioactivity View Larger

Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein (10522-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

SDS-PAGE Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein SDS-PAGE View Larger

2 μg/lane of Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein (10522-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 78-92 kDa and 70-80 kDa, respectively.

Flow Cytometry View Larger

In a functional flow cytometry test, (A) Recombinant SARS-CoV-2 Spike S1 subunit His-tag Protein (Catalog # 10522-CV) binds to HEK293 human embryonic kidney cell line transfected with recombinant human ACE-2 and EGFP. Ligand binding was detected by staining cells with APC-conjugated anti-His Monoclonal Antibody (IC050A), which does not stain the cells in the absence of recombinant protein (B).

Reconstitution Calculator

Reconstitution Calculator

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 S1 Subunit

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 has 65% identity with SARS-CoV-1 S1 subunit,  but only 22% homology with the MERS S1 subunit. 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 S1 subunit 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 Protein, S1 Subunit
Alternate Names
SARS-CoV-2; Spike S1 Subunit

Citations for Recombinant SARS-CoV-2 Spike S1 Subunit His-tag Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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  1. Rapid Generation of Pulmonary Organoids from Induced Pluripotent Stem Cells by Co-Culturing Endodermal and Mesodermal Progenitors for Pulmonary Disease Modelling
    Authors: Mitchell, A;Yu, C;Zhao, X;Pearmain, L;Shah, R;Hanley, KP;Felton, T;Wang, T;
    Biomedicines
    Species: Human
    Sample Types: Organoids
    Applications: Bioassay
  2. A new multiplex SARS-CoV-2 antigen microarray showed correlation of IgG, IgA, and IgM antibodies from patients with COVID-19 disease severity and maintenance of relative IgA and IgM antigen binding over time
    Authors: ML Berre, T Paulov?áko, CM Verissimo, S Doyle, JP Dalton, C Masterson, ER Martínez, L Walsh, C Gormley, JG Laffey, B McNicholas, AJ Simpkin, M Kilcoyne
    PLoS ONE, 2023-03-30;18(3):e0283537.
    Species: Human
    Sample Types: Serum
    Applications: Binding
  3. A new multiplex SARS-CoV-2 antigen microarray showed correlation of IgG, IgA, and IgM antibodies from patients with COVID-19 disease severity and maintenance of relative IgA and IgM antigen binding over time
    Authors: ML Berre, T Paulov?áko, CM Verissimo, S Doyle, JP Dalton, C Masterson, ER Martínez, L Walsh, C Gormley, JG Laffey, B McNicholas, AJ Simpkin, M Kilcoyne
    PLoS ONE, 2023;18(3):e0283537.
    Species: Human
    Sample Types: Serum
    Applications: Binding
  4. Plasmon-enhanced Quantitative Lateral Flow Assay for Femtomolar Detection of SARS-CoV-2 Antibodies and Antigens
    Authors: R Gupta, P Gupta, Z Wang, A Seth, J Morrissey, I George, S Gandra, G Storch, B Parikh, G Genin, S Singamanen
    Research square, 2022-02-21;0(0):.
    Species: Human
    Sample Types: Recombinant Proteins
    Applications: Bioassay

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