Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

Name: Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF
Brand: R&D Systems
SKU: 10778-CV
Price: 524 EUR

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 10778-CV

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D614G, Q677H

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Source:	HEK293
Accession #:	YP_009724390.1

View Full Technical Details for Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

Product Specifications for Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

Source	Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike S1 Subunit protein Val16-Pro681 (Asp614Gly, Gln677His), 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	Protein identity confirmed by mass spectrometry
Predicted Molecular Mass	75 kDa
SDS-PAGE	105-117 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 Examples for Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

	Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein Binding Activity. Recombinant SARS-CoV-2 20C-US Spike S1 Subunit His-tag (Catalog # 10778-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.
	Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein SDS-PAGE. 2 μg/lane of Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag (Catalog # 10778-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 105-117 kDa.
	Binding of ACE-2 to SARS-CoV-2 Spike S1 subunit protein with D614G Q677H mutations by surface plasmon resonance (SPR). Recombinant SARS-CoV-2 Spike protein mutant S1subunit D614G Q677H 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=2.129 nM. (Biacore T200).

Formulation, Preparation and Storage

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.

10778-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.

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Background: Spike S1 Subunit

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). The S1 protein of SARS-CoV-2 shares 65% and 22% amino acid (aa) sequence identity with the S1 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 (6). Based on structural biology studies, the RBD can be oriented either in the up/standing or down/lying state with the up/standing state associated with higher pathogenicity (7). Polyclonal antibodies to the RBD of the SARS-CoV-2 S1 protein have been shown to inhibit interaction with the ACE2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (8). It has been demonstrated that the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (9, 10). 20C.US is a SARS-CoV-2 variant that was first identified late May 2020 and become the most dominant strain in the US. It has not spread to regions outside the US yet. Among other signature point mutations, two of them are found in the S protein (D614G and Q677H) (11).

Item	Value
References	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. Ortega, J.T. et al. (2020) EXCLI J. 19:410. Yuan, Y. et al. (2017) Nat. Commun. 8:15092. Tai, W. et al. (2020) Cell. Mol. Immunol. https://doi.org/10.1016/j.it.2020.03.007. Wang, X. et al. (2020) https://doi.org/10.1038/s41423-020-0424-9. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1. Pater, A. et al. (2021) bioRxiv https://doi.org/10.1101/2021.01.11.426287.
Long Name	Spike Protein, S1 Subunit
Alternate Names	SARS-CoV-2, Spike S1 Subunit

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Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

R&D Systems, Inc. a Bio-Techne Brand

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Product Specifications for Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

Scientific Data Examples for Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: Spike S1 Subunit

Citations for Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

Product Documents for Recombinant SARS-CoV-2 20C-US S1 Subunit His-tag Protein, CF

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