What is the significance of the Arg682Ser, Arg685Ser and Lys986Pro, Val987Pro mutations in Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF (Catalog # 10796-CV)?

Name: Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF
Brand: R&D Systems
SKU: 10796-CV
Rating: 5 (1 reviews)

Furin cleavage resistance is conferred by the Arg682Ser and Arg685Ser mutations. The mutations Lys986Pro and Val987Pro stabilize and promote the prefusion conformation of this Spike protein.

Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

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

Catalog # 10796-CV

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Product Datasheet / COA / SDS

Alpha Variant (UK)

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Key Product Details

Source:	HEK293
Accession #:	YP_009724390.1
Applications:	Bioactivity

View Full Technical Details for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

Product Specifications for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike protein

SARS-CoV-2 B.1.1.7 Spike (Val16-Lys1211)(His69del, Val70del, Tyr145del, Asn501Tyr, Ala570Asp, Asp614Gly, Pro681His, Thr716Iso, Ser982Ala, Asp1118His)(Arg682Ser, Arg685Ser, Lys986Pro, Val987Pro) Accession # YP_009724390.1	GCN4-IZ	6-His tag
N-terminus		C-terminus

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

138 kDa

SDS-PAGE

140-170 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 Examples for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

	Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His-tag Protein Binding Activity. Recombinant SARS-CoV-2 B.1.1.7 Spike (GCN4-IZ) His-tag (Catalog # 10796-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.
	Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His-tag Protein SDS-PAGE. 2 μg/lane of Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His-tag (Catalog # 10796-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 140-170 kDa.
	Binding of ACE-2 to UK variant Spike protein (B.1.1.7) by surface plasmon resonance (SPR). Recombinant SARS-CoV-2 B.1.1.7 (UK Variant) Spike protein (GCN4-IZ) 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 0.046 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=0.6131 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.

10796-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.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 also include MERS-CoV 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). 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 S Protein of the SARS-CoV-2 virus, like the SARS-CoV-1 counterpart, binds a metallopeptidase, Angiotensin-Converting Enzyme 2 (ACE-2), but with much higher affinity and faster binding kinetics through the receptor binding domain (RBD) located in the C-terminal region of S1 subunit (6). It has been demonstrated that the S Protein can invade host cells through the CD147/EMMPRIN receptor and mediate membrane fusion (7, 8). Polyclonal antibodies to the RBD of the SARS-CoV-2 protein have been shown to inhibit interaction with the ACE-2 receptor, confirming RBD as an attractive target for vaccinations or antiviral therapy (9). There is also promising work showing that the RBD may be used to detect presence of neutralizing antibodies present in a patient's bloodstream, consistent with developed immunity after exposure to the SARS-CoV-2 (10). Several emerging SARS-CoV-2 genomes have been identified including the B 1.1.7 (United Kingdom) variant (11). The B 1.1.7 variant contains 1 significant mutation of interest in the RBD domain, N501Y, which has been shown to result in enhanced binding affinity for hACE-2 (12). Further, the B 1.1.7 variant appears to more easily transmissible, exhibit increased viral loads and, potentially, be associated with higher mortality rates compared to preexisting variants (11, 13).

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. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1. Isabel, et al. (2020) Sci Rep 10, 14031. https://doi.org/10.1038/s41598-020-70827-z. Tai, W. et al. (2020) Cell. Mol. Immunol. https://doi.org/10.1016/j.it.2020.03.007.1. Okba, N.M.A. et al. (2020) Emerg. Infect. Dis. https://doi.org/10.3201/eid2607.200841. Kidd, M. et al. (2021) The Journal of Infectious Diseases https://doi.org/10.1093/infdis/jiab082. Zahradník, J. et al. (2021) bioRxiv https://doi.org/10.1101/2021.01.06.425392. Davies, N.G. (2020) medRxiv doi:10.1101/2020.12.24.20248822.
Long Name	Spike Protein
Entrez Gene IDs	918758 (HCoV-229E); 2943499 (HCoV-NL63); 39105218 (HCoV-OC43); 37616432 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)
Alternate Names	2019-nCoV S Protein, 2019-nCoV Spike, COVID-19 Spike, E2, Human coronavirus spike glycoprotein, Peplomer protein, S Protein, S glycoprotein, SARS-COV-2 S protein, SARS-COV-2 Spike glycoprotein, SARS-CoV-2, SARSCOV2 Spike protein, Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein, Spike, Spike glycoprotein, surface glycoprotein

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Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay

A Shajahan et al. (2022), Site Specific N- and O-glycosylation mapping of the Spike Proteins of SARS-CoV-2 Variants of Concern Research square, 0(0).
PMID: 36415454

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Application: Immunoassay Standard

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FAQs for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

What is the significance of the Arg682Ser, Arg685Ser and Lys986Pro, Val987Pro mutations in Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF (Catalog # 10796-CV)?

Furin cleavage resistance is conferred by the Arg682Ser and Arg685Ser mutations. The mutations Lys986Pro and Val987Pro stabilize and promote the prefusion conformation of this Spike protein.

Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

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

Key Product Details

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Product Specifications for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

Scientific Data Examples for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: Spike

Citations for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF (1)

Customer Reviews for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF (1)

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Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

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Product Documents for Recombinant SARS-CoV-2 B.1.1.7 Spike GCN4-IZ His Protein, CF

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