Recombinant SARS-CoV-2 E484K N501Y Spike RBD His Protein, CF

Catalog # 10788-CV | R&D Systems, Inc. a Bio-Techne Brand
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Key Product Details

Accession #

YP_009724390.1

Source

HEK293

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike RBD protein
Arg319-Phe541 (Glu484Lys, Asn501Tyr), with a C-terminal 6-His tag

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

Arg319

Predicted Molecular Mass

26 kDa

SDS-PAGE

32-40 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 E484K N501Y Spike RBD His Protein, CF

Recombinant SARS-CoV-2 E484K N501Y Spike RBD His-tag Protein Binding Activity.

Recombinant SARS-CoV-2 E484K N501Y Spike RBD His-tag (Catalog # 10788-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

Recombinant SARS-CoV-2 E484K N501Y Spike RBD His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant SARS-CoV-2 E484L N501Y Spike RBD His-tag (Catalog # 10788-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 32-40 kDa.

Formulation, Preparation and Storage

10788-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.

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 also include MERS 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). A metallopeptidase, angiotensin-converting enzyme 2 (ACE2), has been identified as a functional receptor for SARS-CoV-2 through interaction with a receptor binding domain (RBD) located at the C-terminus of S1 subunit (6,7). The RBD of SARS-CoV-2 shares 73% amino acid (aa) identity with the RBD of the SARS-CoV-1, but only 22% aa identity with the RBD of MERS. A SARS-CoV-2 variant carrying the aa substitution N501Y in the RBD is one of the most prevalent mutations found Covid-19 cases (8-10). This mutation was first identified in the virus variant (B.1.1.7 lineage) originally found in London and the southeast UK but rapidly spread globally (8,9). This new virus variant was reported 56% more transmissible than other preexisting variants (11). Another mutation E484K, in addition to the N501Y mutation, was also later identified in variants found in South Arica (B.1.351 lineage) and Brazil (P.1 lineage). Although there is no evidence to date that these two aa mutations in RBD causes more severe illness, whether such mutations would decrease the efficacy of vaccine-induced immunity is still under investigation.

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. Li, W. et al. (2003) Nature 426:450.
  7. Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
  8. Kozlov, Max (2020) TheScientist https://www.the-scientist.com/news-opinion/new-sars-cov-2-variant-spreading-rapidly-in-uk-68292.
  9. Wise, J. (2020) B.M.J 371:m4857.
  10. Tang, J.W. et al. (2020) J. Infect. doi: 10.1016/j.jinf.2020.12.024.
  11. Davies, N.G. (2020) medRxiv doi:10.1101/2020.12.24.20248822.

Long Name

Spike Receptor Binding Domain

Alternate Names

Spike RBD

Entrez Gene IDs

3200426 (HCoV-HKU1); 14254594 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)

Gene Symbol

S

UniProt

YP_009724390.1

Product Documents for Recombinant SARS-CoV-2 E484K N501Y Spike RBD His Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant SARS-CoV-2 E484K N501Y Spike RBD His Protein, CF

For research use only

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