Recombinant SARS-CoV-2 Spike His Protein, CF

Ectodomain, Stabilized Prefusion Conformation, Resistant to Furin Cleavage
Catalog # Availability Size / Price Qty
10549-CV-100
10549-CV-01M
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Graph showing SARS-CoV-2 Spike protein binding to ACE-2
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Product Details
Citations (12)
FAQs
Reviews (2)

Recombinant SARS-CoV-2 Spike His Protein, CF Summary

Why choose R&D Systems SARS-CoV-2 Spike Protein?

  • Val16-Lys1211 with a C-term His tag.​
  • Stabilizing mutations K986P and V987P promote the prefusion conformation.
  • Two mutations, R682S and R685S, eliminate a Furin protease cleavage site.
  • Guaranteed Bioactivity and High Purity: Bioactivity tested by functional ELISA and purity determined by SDS-PAGE to be greater than 95%.
  • Lot-to-Lot Consistency: Stringent QC testing performed on each lot to ensure consistent activity and purity.
  • Bulk Quantities Available: Bulk up and save with large mass quantities to meet your research needs. Supply agreements available, partner with us. Please contact us.
  • Most Respected, Most Cited Brand in Proteins: With over 35 years of providing the best recombinant proteins to the scientific community, R&D Systems continues to lead the industry in quality, activity, and purity.

Interested in other Coronavirus proteins including SARS-CoV-2 Spike proteins? View Products

Product Specifications

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.
Activity
Measured by its binding ability in a functional ELISA with Recombinant Human ACE-2 His-tag  (Catalog # 933-ZN).
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
Accession #
N-terminal Sequence
Analysis
Val16
Predicted Molecular Mass
134 kDa
SDS-PAGE
144-175 kDa, under reducing conditions

Product Datasheets

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10549-CV

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.

Scientific Data

Bioactivity Graph showing SARS-CoV-2 Spike protein binding to ACE-2 View Larger

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).

SDS-PAGE SDS-PAGE gel of purified recombinant SARS-CoV-2 Spike protein View Larger

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.

Surface Plasmon Resonance (SPR) Surface plasmon resonance sensorgram of SARS-CoV-2 Spike protein binding to Human ACE-2 View Larger

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).

SDS-PAGE SDS-PAGE gel of purified recombinant SARS-CoV-2 Spike protein run side-by-side against a competitor's Spike protein View Larger

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.

Flow Cytometry Flow cytometry scatter plot showing SARS-CoV-2 Spike protein binds to ACE-2 expressing HEK293 cells View Larger

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).

Reconstitution Calculator

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 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).

References
  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. Ortega, J.T. et al. (2020) EXCLI J. 19:410.
  7. Yuan, Y. et al. (2017) Nat. Commun. 8:15092.
  8. Tai, W. et al. (2020) Cell. Mol. Immunol. https://doi.org/10.1016/j.it.2020.03.007.
  9. Wang, X. et al. (2020) https://doi.org/10.1038/s41423-020-0424-9.
  10. Wang, K. et al. (2020) bioRxiv https://www.biorxiv.org/content/10.1101/2020.03.14.988345v1.
  11. Korber, B. et al. (2020) Cell 182, 812.
  12. 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 glycoprotein; S Protein; SARS-COV-2 S protein; SARS-COV-2 Spike glycoprotein; SARSCOV2 Spike protein; SARS-CoV-2; Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein; Spike glycoprotein; Spike; surface glycoprotein

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

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

12 Citations: Showing 1 - 10
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  1. Suppression of host humoral immunity by Borrelia burgdorferi varies over the course of infection
    Authors: Williams, MT;Zhang, Y;Pulse, ME;Berg, RE;Allen, MS;
    Infection and immunity
    Species: Murine polyomavirus strain A3
    Sample Types: Recombinant Protein, Serum, In Vivo
    Applications: In vivo assay, ELISA Standard
  2. Filamentous fungus-produced human monoclonal antibody provides protection against SARS-CoV-2 in hamster and non-human primate models
    Authors: Kaiser, FK;Hernandez, MG;Krüger, N;Englund, E;Du, W;Mykytyn, AZ;Raadsen, MP;Lamers, MM;Rodrigues Ianiski, F;Shamorkina, TM;Snijder, J;Armando, F;Beythien, G;Ciurkiewicz, M;Schreiner, T;Gruber-Dujardin, E;Bleyer, M;Batura, O;Erffmeier, L;Hinkel, R;Rocha, C;Mirolo, M;Drabek, D;Bosch, BJ;Emalfarb, M;Valbuena, N;Tchelet, R;Baumgärtner, W;Saloheimo, M;Pöhlmann, S;Grosveld, F;Haagmans, BL;Osterhaus, ADME;
    Nature communications
    Species: Human hepegivirus
    Sample Types: Whole Cells
    Applications: ELISA Capture
  3. Evidence, detailed characterization and clinical context of complement activation in acute multisystem inflammatory syndrome in children
    Authors: G Sinkovits, J Schnur, L Hurler, P Kiszel, ZZ Prohászka, P Sík, E Kajdácsi, L Cervenak, V Maráczi, M Dávid, B Zsigmond, É Rimanóczy, C Bereczki, L Willems, EJM Toonen, Z Prohászka
    Scientific Reports, 2022-11-17;12(1):19759.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: ELISA Standard
  4. An experimental test of the nicotinic hypothesis of COVID-19
    Authors: NE Godellas, GD Cymes, C Grosman
    Proceedings of the National Academy of Sciences of the United States of America, 2022-10-24;119(44):e2204242119.
    Species: Human
    Sample Types: Protein
    Applications: Control
  5. Calcium dobesilate reduces SARS-CoV-2 entry into endothelial cells by inhibiting virus binding to heparan sulfate
    Authors: Y Kiyan, A Schultalbe, E Chernobriv, S Tkachuk, S Rong, N Shushakova, H Haller
    Scientific Reports, 2022-10-07;12(1):16878.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  6. Waning of SARS-CoV-2 Seropositivity among Healthy Young Adults over Seven Months
    Authors: CS Lea, K Simeonsson, AM Kipp, C McNeill, L Wilcox, W Irish, H Morris, OM Diaz, JT Fallon, RL Roper
    Vaccines, 2022-09-15;10(9):.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Capture
  7. SARS-CoV-2 Spike Proteins and Cell-Cell Communication Inhibits TFPI and Induces Thrombogenic Factors in Human Lung Microvascular Endothelial Cells and Neutrophils: Implications for COVID-19 Coagulopathy Pathogenesis
    Authors: B Bhargavan, GD Kanmogne
    International Journal of Molecular Sciences, 2022-09-09;23(18):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  8. Potent universal beta-coronavirus therapeutic activity mediated by direct respiratory administration of a Spike S2 domain-specific human neutralizing monoclonal antibody
    Authors: MS Piepenbrin, JG Park, A Deshpande, A Loos, C Ye, M Basu, S Sarkar, AM Khalil, D Chauvin, J Woo, P Lovalenti, NB Erdmann, PA Goepfert, VL Truong, RA Bowen, MR Walter, L Martinez-S, JJ Kobie
    PloS Pathogens, 2022-07-21;18(7):e1010691.
    Species: Primate - C. aethiops
    Sample Types: Whole Cells
    Applications: Surface Plasmon Resonance
  9. Cross-reactive antibodies elicited to conserved epitopes on SARS-CoV-2 spike protein after infection and vaccination
    Authors: ES Geanes, C LeMaster, ER Fraley, S Khanal, R McLennan, E Grundberg, R Selvaranga, T Bradley
    Scientific Reports, 2022-04-20;12(1):6496.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Capture
  10. Humoral Immunogenicity and Reactogenicity of the Standard ChAdOx1 nCoV-19 Vaccination in Taiwan
    Authors: JH Chang, JF Chiou, CS Hung, MC Liu, HW Chang, SY Hong, CY Wang, YL Lin, YC Hsieh, CL Chung, YS Su, SS Hsiao, D Liu, JJ Liang, CC Liao, CS Chang, KS Lai, HC Chuang, KL Chien, WC Wu, YG Lee, SE Lin, YK Shen, CF Hsu, JC Wang, SH Hsiao
    Vaccines, 2022-02-17;10(2):.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Capture
  11. The spike protein of SARS-CoV-2 induces endothelial inflammation through integrin alpha5beta1 and NF-kappaB signaling
    Authors: JP Robles, M Zamora, E Adan-Castr, L Siqueiros-, G Martinez d, C Clapp
    The Journal of Biological Chemistry, 2022-02-07;0(0):101695.
    Species: Human, Mouse, Rat
    Sample Types: In Vivo, Recombinant Protein
    Applications: Bioassay, In Vivo
  12. Beneficial Effects of Mineralocorticoid Receptor Pathway Blockade against Endothelial Inflammation Induced by SARS-CoV-2 Spike Protein
    Authors: E Jover, L Matilla, M Garaikoetx, A Fernández-, P Muntendam, F Jaisser, P Rossignol, N López-Andr
    Biomedicines, 2021-06-03;9(6):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

FAQs

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

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Recombinant SARS-CoV-2 Spike (Active Trimer) His Protein, CF
By Eva Piano Mortari on 02/15/2021
Application: FACS

Recombinant SARS-CoV-2 Spike (Active Trimer) His Protein, CF
By Anonymous on 10/09/2020
Application: Immunoassay Standard