Recombinant SARS-CoV-2 B.1.526 Spike GCN4-IZ His Protein, CF

R&D Systems | Catalog # 10848-CV

E484K Point Mutation. Iota Variant
R&D Systems
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Key Product Details

  • R&D Systems HEK293-derived Recombinant SARS-CoV-2 B.1.526 Spike GCN4-IZ His Protein (10848-CV)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

HEK293

Accession Number

Applications

Bioactivity
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Product Specifications

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike protein
SARS-CoV-2 Spike
(Val16-Lys1211)(Thr95Ile, Asp253Gly, Glu484Lys, Asp614Gly, Ala701Val)(Arg682Ser, Arg685Ser, Lyso986Pro, Val987Pro)
Accession # YP_009724390.1
GCN4-IZ 6-His tag
N-terminus C-terminus

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Val16

Predicted Molecular Mass

138 kDa

SDS-PAGE

145-165 kDa, under reducing conditions.

Activity

Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag (Catalog # 933-ZN).

Scientific Data Images for Recombinant SARS-CoV-2 B.1.526 Spike GCN4-IZ His Protein, CF

Recombinant SARS-CoV-2 B.1.526 Spike (GCN4-IZ) His-tag Protein Binding Activity.

Recombinant SARS-CoV-2 B.1.526 Spike (GCN4-IZ) His-tag (Catalog # 10848-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

Recombinant SARS-CoV-2 B.1.526 Spike (GCN4-IZ) His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant SARS-CoV-2 B.1.526 Spike (GCN4-IZ) His-tag Protein (Catalog # 10848-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 145-165 kDa.

Formulation, Preparation, and Storage

10848-CV
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 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.

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

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). The S protein of SARS-CoV-2 shares 75% and 29% amino acid sequence identity with S protein of SARS-CoV-1 and MERS, respectively. The S Protein of the SARS‑CoV‑2 virus, like the SARS-CoV-1 counterpart, binds a metallopeptidase, Angiotensin-Converting Enzyme 2 (ACE-2), but with much higher affinity and faster binding kinetics through the receptor binding domain (RBD) located in the C-terminal region of S1 subunit (6). It has been demonstrated that the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (7, 8). 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 (9). 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 (10). Several emerging SARS-CoV-2 genomes have been identified including the B.1.526 variant, with an additional B.1.526-E484K variant identified in New York City in late 2020 (11). The E484K mutation is located in the RBD and is also present in the B.1.351 (South Africa) and P.1 (Brazil) variants (12). Several studies suggest E484K as a potentially crucial mutation as it creates a new site for hACE-2 binding and enhanced binding affinity, has been shown to confer resistance to several monoclonal antibodies and is responsible for the first confirmed SARS-CoV-2 reinfection (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. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
  7. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1.
  8. Isabel, et al. (2020) Sci Rep 10, 14031. https://doi.org/10.1038/s41598-020-70827-z.
  9. Tai, W. et al. (2020) Cell. Mol. Immunol. https://doi.org/10.1016/j.it.2020.03.007.1.
  10. Okba, N.M.A. et al. (2020) Emerg. Infect. Dis. https://doi.org/10.3201/eid2607.200841.
  11. Lasek-Nesselquist, E. et al. (2021) medrxiv https://doi.org/10.1101/2021.02.26.21251868.
  12. Ferrareze, P.A.G. et al. (2021) bioRxiv https://doi.org/10.1101/2021.01.27.42689.
  13. Wang, W.B. et al. (2021) bioRxiv https://doi.org/10.1101/2021.02.17.431566.
  14. Vasques Nonaka, C.K. et al. (2021) Emerg Infect Dis. https://doi.org/10.3201/eid2705.210191.

Long Name

Spike Protein

Alternate Names

S Protein

Entrez Gene IDs

918758 (HCoV-229E); 2943499 (HCoV-NL63); 39105218 (HCoV-OC43); 37616432 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)

Gene Symbol

S

UniProt

Additional Spike Products

Product Documents for Recombinant SARS-CoV-2 B.1.526 Spike GCN4-IZ His Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant SARS-CoV-2 B.1.526 Spike GCN4-IZ His Protein, CF

For research use only

Related Research Areas

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