Recombinant SARS-CoV-2 B.1.617.2 Spike RBD His Protein, CF
Recombinant SARS-CoV-2 B.1.617.2 Spike RBD His Protein, CF Summary
Arg319-Phe541 (Leu452Arg, Thr478Lys), a C-terminal 6-His tag
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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.
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.
|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.
Recombinant SARS-CoV-2 B.1.617.2 Spike RBD (Catalog # 10876-CV) binds Recombinant Human ACE-2 (933-ZN) in a functional ELISA.
2 μg/lane of Recombinant SARS-CoV-2 B.1.617.2 Spike RBD His-tag Protein (Catalog # 10876-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 30-37 kDa.
Recombinant SARS-CoV-2 B.1.617.2 Delta variant Spike protein RBD His-tag (Catalog # 10876-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 94.3 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=3.66 nM. (Biacore T200).
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 (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). Based on amino acid (aa) sequence homology, the SARS-CoV-2 S1 subunit RBD has 73% identity with the RBD of the SARS-CoV-1 S1 RBD, but only 22% homology with the MERS S1 RBD. 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).
- Wu, F. et al. (2020) Nature 579:265.
- Tortorici, M.A. and D. Veesler (2019) Adv. Virus Res. 105:93.
- Bosch, B.J. et al. (2003) J. Virol. 77:8801.
- Belouzard, S. et al. (2009) Proc. Natl. Acad. Sci. 106:5871.
- Millet, J.K. and G.R. Whittaker (2015) Virus Res. 202:120.
- Li, W. et al. (2003) Nature 426:450.
- Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
- Yadav, P.D. et al. (2021) bioRxiv https://doi.org/10.1101/2021.04.23.441101.
- Cherian, S. et al. (2021) bioRxiv https://doi.org/10.1101/2021.04.22.440932.
- Bernal, J. et al. (2021) medRxiv https://doi.org/10.1101/2021.05.22.21257658.
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