Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD His Protein

N501Y Mutation found in UK, South African, and Brazilian Variants
Catalog # Availability Size / Price Qty
10730-CV-100
Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD His-tag Protein Binding Activity.
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Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD His Protein Summary

Product Specifications

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.
Activity
Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag  (Catalog # 933-ZN).
Source
Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike RBD protein
Arg319-Phe541 (Asn501Tyr), with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Arg319
Predicted Molecular Mass
26 kDa
SDS-PAGE
34-38 kDa, under reducing conditions

Product Datasheets

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.

10730-CV

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 250 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.

Data Images

Binding Activity Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD His-tag Protein Binding Activity. View Larger

Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD His-tag (Catalog # 10730-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

SDS-PAGE Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD His-tag Protein SDS-PAGE. View Larger

2 μg/lane of Recombinant SARS-CoV-2 B.1.1.7 N501Y Spike RBD His-tag (Catalog # 10730-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 34-38 kDa.

Surface Plasmon Resonance (SPR) Binding of ACE-2 to SARS-CoV-2 Spike RBD N501Y by surface plasmon resonance (SPR). View Larger

Recombinant SARS-CoV-2 Spike RBD N501Y His-tag was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (933-ZN) was measured at a concentration range between 0.18 nM and 47.2 nM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of KD=0.3845 nM. (Biacore T200).

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 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% aa identity with the RBD of the SARS-CoV-1, but only 22% amino acid (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 in 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). The N501Y mutation was also later identified in variants found in South Africa (B.1.351 lineage) and Brazil (P.1 lineage). Although there is no evidence to date that B.1.1.7 causes more severe illness, whether the N501Y mutation in RBD 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
Entrez Gene IDs
3200426 (HCoV-HKU1); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)
Alternate Names
2019-nCoV Peplomer protein; 2019-nCoV S Protein; 2019-nCoV Spike; COVID-19 Spike; E2; Human coronavirus spike glycoprotein; Peplomer protein; S glycoprotein; S protein; SARS-COV-2 E2; SARS-COV-2 Peplomer protein; SARS-COV-2 S protein; SARS-COV-2 Spike glycoprotein; SARSCOV2 Spike protein; SARS-CoV-2; Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein; Spike glycoprotein; Spike RBD; Spike S1 RBD; Spike S1; Spike; surface glycoprotein

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