Recombinant SARS-CoV-2 Spike His Protein, CF

Name: Recombinant SARS-CoV-2 Spike His Protein, CF
Brand: R&D Systems
SKU: 10549-CV
Rating: 5 (2 reviews)

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

Catalog # 10549-CV

Citations (9)

(2)

Product Datasheet / COA / SDS

Ectodomain, Stabilized Prefusion Conformation, Resistant to Furin Cleavage

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Key Product Details

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

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Product Specifications for Recombinant SARS-CoV-2 Spike His Protein, CF

Source	Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike protein Val16-Lys1211 (Arg682Ser, Arg685Ser, Lys986Pro and Val987Pro) with a C-terminal 6-His tag Suitable for use in serological assay development
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	134 kDa
SDS-PAGE	144-175 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 Spike His Protein, CF

	Recombinant SARS-CoV-2 Spike His Protein Binding Activity R&D Systems Recombinant SARS-CoV-2 Spike (10549-CV) binds Recombinant Human ACE-2 (933-ZN) in a functional ELISA. The binding activity is approximately 4-fold greater than a top competitor’s Spike protein (full ectodomain).
	Recombinant SARS-CoV-2 Spike His Protein SDS-PAGE 2 μg/lane of Recombinant SARS-CoV-2 Spike His Protein (10549-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 144-175 kDa.
	Binding of ACE-2 to SARS-CoV-2 Spike by surface plasmon resonance (SPR). Recombinant SARS-CoV-2 Spike His Protein (Catalog # 10549-CV) was immobilized on a Biacore Sensor Chip CM5, and binding to recombinant human ACE-2 (Catalog # 933-ZN) was measured at a concentration range between 0.37 nM and 93.5 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=1.720 nM. (Biacore T200).
	Recombinant SARS-CoV-2 Spike His Protein SDS-PAGE SDS-PAGE of 1 µg of R&D Systems Recombinant SARS-CoV-2 Spike (10549-CV) or a competitor’s Spike protein were run under reducing or non-reducing conditions and visualized by silver staining. The R&D Systems Spike protein runs as a single band compared to the competition.
	Detection of SARS-CoV-2 Spike Protein bound to ACE-2 expressing cells by flow cytometry. In a functional flow cytometry test, (A) Recombinant SARS-CoV-2 Spike His-tag Protein (Catalog # 10549-CV) binds to HEK293 human embryonic kidney cell line transfected with recombinant human ACE-2 and EGFP. Ligand binding was detected by staining cells with APC-conjugated anti-His Monoclonal Antibody (IC050A), which does not stain the cells in the absence of recombinant protein (B).

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.

10549-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

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 S protein of SARS-CoV-2 shares 75% and 29% amino acid (aa) sequence identity with the 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 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 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). A SARS-CoV-2 variant carrying the S protein aa change D614G has become the most prevalent form in the global pandemic and has been associated with greater infectivity and higher viral load (11, 12).

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. Korber, B. et al. (2020) Cell 182, 812. Zhang, L. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.06.12.148726v1.
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

Citations for Recombinant SARS-CoV-2 Spike His Protein, CF (9)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant SARS-CoV-2 Spike His Protein, CF include:

Species: Human
Sample Types: Serum
Applications: ELISA Capture

JH Chang et al. (2022), Humoral Immunogenicity and Reactogenicity of the Standard ChAdOx1 nCoV-19 Vaccination in Taiwan Vaccines, 10(2).
PMID: 35214770
Species: Human
Sample Types: Serum/Plasma
Applications: ELISA

ES Geanes et al. (2022), Cross-reactive antibodies elicited to conserved epitopes on SARS-CoV-2 spike protein after infection and vaccination Scientific Reports, 12(1):6496.
PMID: 35444221
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay

JP Robles et al. (2022), The spike protein of SARS-CoV-2 induces endothelial inflammation through integrin alpha5beta1 and NF-kappaB signaling The Journal of Biological Chemistry, 0(0):101695.
PMID: 35143839
Species: Human
Sample Types: Serum
Applications: ELISA Capture

CS Lea et al. (2022), Waning of SARS-CoV-2 Seropositivity among Healthy Young Adults over Seven Months Vaccines, 10(9).
PMID: 36146610
Species: Primate - C. aethiops
Sample Types: Whole Cells
Applications: Surface Plasmon Resonance

MS Piepenbrin et al. (2022), Potent universal beta-coronavirus therapeutic activity mediated by direct respiratory administration of a Spike S2 domain-specific human neutralizing monoclonal antibody PloS Pathogens, 18(7):e1010691.
PMID: 35862475
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay

Y Kiyan et al. (2022), Calcium dobesilate reduces SARS-CoV-2 entry into endothelial cells by inhibiting virus binding to heparan sulfate Scientific Reports, 12(1):16878.
PMID: 36207386
Species: Human
Sample Types: Recombinant Protein
Applications: ELISA Standard

G Sinkovits et al. (2022), Evidence, detailed characterization and clinical context of complement activation in acute multisystem inflammatory syndrome in children Scientific Reports, 12(1):19759.
PMID: 36396679
Species: Human
Sample Types: Protein
Applications: Control

NE Godellas et al. (2022), An experimental test of the nicotinic hypothesis of COVID-19 Proceedings of the National Academy of Sciences of the United States of America, 119(44):e2204242119.
PMID: 36279466
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

E Jover et al. (2021), Beneficial Effects of Mineralocorticoid Receptor Pathway Blockade against Endothelial Inflammation Induced by SARS-CoV-2 Spike Protein Biomedicines, 9(6).
PMID: 34204890

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Customer Reviews for Recombinant SARS-CoV-2 Spike His Protein, CF (2)

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Recombinant SARS-CoV-2 Spike (Active Trimer) His Protein, CF

Name: Eva Piano Mortari

Application: FACS

Verified Customer | Posted 02/15/2021
Recombinant SARS-CoV-2 Spike (Active Trimer) His Protein, CF

Name: Anonymous

Application: Immunoassay Standard

Verified Customer | Posted 10/09/2020

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