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Recombinant SARS-CoV-2 D614G Spike His-tag, Protein CF

Bio-Techne includes R&D Systems | Catalog # 10620-CV

CHO Expressed His-tag. Mutations: D614G, R682S, R685S, K986P, V987P
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10620-CV-100

Key Product Details

Source

CHO

Accession #

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived sars-cov-2 Spike protein
Val16-Lys1211 (Asp614Gly, Arg682Ser, Arg685Ser, Lys986Pro, Val987Pro), 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

Val16

Predicted Molecular Mass

134 kDa

SDS-PAGE

150-170 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 D614G Spike His-tag, Protein CF

Recombinant SARS-CoV-2 D614 G Spike His-tag, Protein Binding Activity.

Recombinant SARS-CoV-2 D614G Spike His-tag (Catalog # 10620-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

Recombinant SARS-CoV-2 D614G Spike His-tag, Protein SDS-PAGE.

2 μg/lane of Recombinant SARS-CoV-2 D614G Spike His-tag, Protein (Catalog # 10620-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 150-170 kDa.
Surface plasmon resonance (SPR) sensorgram of Human ACE-2 binding to SARS-CoV-2 Spike protein, D614G mutant

Binding of ACE-2 to SARS-CoV-2 Spike protein, D614G mutant by surface plasmon resonance (SPR).

Recombinant SARS-CoV-2 Spike protein D614G 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 0.092 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=2.099 nM. (Biacore T200).

Formulation, Preparation and Storage

10620-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

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 the 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 SARS-CoV-2 variant carrying the S protein amino acid (aa) change D614G has become the most prevalent form in the global pandemic and has been associated with greater infectivity and higher viral load (6,7). The S protein of SARS-CoV-2 shares 75% and 29% aa 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 Angiotensin-Converting Enzyme 2 (ACE2), but with much higher affinity and faster binding kinetics through the receptor binding domain (RBD) located in the C-terminal region of S1 (8). It has been demonstrated that the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (9, 10). It is proposed that the D614G mutation introduces an additional elastase cleavage site near the S1-S2 junction and this additional processing helps with viral entry (11).

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. Korber, B. et al. (2020) Cell 182:812.
  7. Zhang, L. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.06.12.148726v1.
  8. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
  9. Wang, X. et al. (2020) https://doi.org/10.1038/s41423-020-0424-9.
  10. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1.
  11. Bhattacharyya, C. et al. (2020) bioRxiv https://doi.org/10.1101/2020.05.04.075911.

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 D614G Spike His-tag, Protein CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant SARS-CoV-2 D614G Spike His-tag, Protein CF

For research use only

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