Recombinant SARS-CoV-2 B.1.617.1 RBD His Avi-tag Protein, CF

Biotinylated Recombinant SARS-CoV-2 B.1.617.1 Spike RBD His-tag Avi-tag (Catalog # AVI10846) binds Recombinant Human ACE-2 Fc Chimera (Catalog # 10544-ZN) in a functional ELISA.

2 μg/lane of Biotinylated Recombinant SARS-CoV-2 B.1.617.1 Spike RBD His-tag Avi-tag Protein (Catalog # AVI10846) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 31-38 kDa.

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

Recombinant SARS-CoV-2 B.1.617.1 RBD His Avi-tag Protein, CF Summary

• Additional Information: Kappa Variant L452R E484Q
• Details of Functionality: Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 Fc Chimera (Catalog # 10544-ZN).
• Source: Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike RBD protein
  

  SARS-CoV-2 B.1.617.1 Spike RBD (Arg319-Phe541) (Leu452Arg, Glu484Gln) Accession # YP_009724390.1	6-His tag	Avi-tag
  N-terminus		C-terminus

• Accession #: YP_009724390.1
• N-terminal Sequence: Arg319
• Structure / Form: Biotinylated via Avi-tag
• 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: 26 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: 35-40 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.1.617.1 RBD His Avi-tag Protein, CF

• Spike RBD

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 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 (B.1.617.1) carrying the aa substitution L452R and E484Q in the RBD was identified as a prevalent strain in India (8, 9). Whether these mutations in RBD would cause more severe symptom or decrease the efficacy of vaccine-induced immunity is still under investigation. Our Avi-tag Biotinylated SARS-CoV-2 B.1.617.1 Spike RBD features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of bionylation and the rest of the protein is unchanged so there is no interference in the protein bioactivity.

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.

Bioinformatics

• Uniprot:
  • YP_009724390.1(Rabbit)