Recombinant SARS-CoV-2 B.1.617.2 Spike GCN4-IZ Protein, CF

Recombinant SARS-CoV-2 Spike B.1.617.2 (GCN4-IZ) His-tag (Catalog # 10878-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

2 μg/lane of Recombinant SARS-CoV-2 B.1.617.2 Spike His-tag Protein (Catalog # 10878-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 144-170 kDa.

Recombinant SARS-CoV-2 B.1.617.2 Delta variant Spike protein His-tag (Catalog #10878-CV) was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (933-ZN) was measured at a concentration range between 0.046 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=1.29 nM.

• Reactivity: V
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant SARS-CoV-2 B.1.617.2 Spike GCN4-IZ Protein, CF Summary

• Additional Information: Delta Variant (India) His-tag
• Details of Functionality: 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 protein
  

  SARS-CoV-2 Spike (Val16-Lys1211)(Thr19Arg, Gly142Asp, Glu156Gly, Phe157del, Arg158del, Leu452Arg, Thr478Lys, Asp614Gly, Pro681Arg, Asp950Asn)(Arg682Ser, Arg685Ser, Lys986Pro, Val987Pro) Accession # YP_009724390.1	GCN4-IZ	6-His tag
  N-terminus		C-terminus

• Accession #: YP_009724390.1
• N-terminal Sequence: Val16
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 138 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 144-170 kDa, under reducing conditions.

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant SARS-CoV-2 B.1.617.2 Spike GCN4-IZ Protein, CF

• 2019-nCoV S Protein
• 2019-nCoV Spike
• COVID-19 Spike
• E2
• Human coronavirus spike glycoprotein
• Peplomer protein
• S glycoprotein
• S 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
• surface glycoprotein

Background

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 (ACE-2), 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 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 SARS-CoV-2 delta variant (B.1.617.2) carrying the amino acid substitution L452R and T478K in the RBD was identified as a prevalent strain in India and has been detected in more than 40 countries (8, 9). It has higher transmissible rate and more resistant to vaccine (10).

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. Yadav, P.D. et al. (2021) bioRxiv https://doi.org/10.1101/2021.04.23.441101.
9. Cherian, S. et al. (2021) bioRxiv https://doi.org/10.1101/2021.04.22.440932.
10. Bernal, J. et al. (2021) medRxiv https://doi.org/10.1101/2021.05.22.21257658.

Publications for SARS-CoV-2 Spike (10878-CV)(4)

Publications using 10878-CV

• Shajahan, A;Pepi, LE;Kumar, B;Murray, NB;Azadi, P; Site specific N- and O-glycosylation mapping of the spike proteins of SARS-CoV-2 variants of concern Scientific reports 2023-06-21 [PMID: 37344512]
• Harless, WW;Lewis, B;Qorri, B;Abdulkhalek, S;Szewczuk, MR; Novel Therapeutic Target Critical for SARS-CoV-2 Infectivity and Induction of the Cytokine Release Syndrome Cells 2023-05-07 [PMID: 37174732] (Bioassay, N/A)
• A Shajahan, L Pepi, B Kumar, N Murray, P Azadi Site Specific N- and O-glycosylation mapping of the Spike Proteins of SARS-CoV-2 Variants of Concern Research square, 2022-11-16;0(0):. 2022-11-16 [PMID: 36415454] (Bioassay, Human)
• B Bhargavan, GD Kanmogne SARS-CoV-2 Spike Proteins and Cell-Cell Communication Inhibits TFPI and Induces Thrombogenic Factors in Human Lung Microvascular Endothelial Cells and Neutrophils: Implications for COVID-19 Coagulopathy Pathogenesis International Journal of Molecular Sciences, 2022-09-09;23(18):. 2022-09-09 [PMID: 36142345] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • YP_009724390.1(Llama)