Recombinant SARS-CoV-2 S GCN4-IZ Alexa Fluor® 488 Protein

R&D Systems | Catalog # AFG10786

B.1.351 His-tag Beta Variant (South Africa)
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Key Product Details

  • R&D Systems HEK293-derived Recombinant SARS-CoV-2 S GCN4-IZ Alexa Fluor® 488 Protein (AFG10786)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

HEK293

Accession Number

YP_009724390.1

Structure / Form

Labeled with Alexa Fluor® 488 via amines


Excitation Wavelength: 488 nm  
Emission Wavelength: 515­-545 nm

Conjugate

Alexa Fluor 488 (Excitation= 488 nm, Emission= 515-545 nm)

Applications

Bioactivity
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Product Specifications

Source

Human embryonic kidney cell, HEK293-derived sars-cov-2 Spike protein
SARS-CoV-2 B.1.351 Spike
(Val16-Lys1211) (Asp80Ala, Asp215Gly, Leu242 del, Ala243 del, Leu244 del, Lys417Asn, Glu484Lys, Asn501Tyr, Asp614Gly, Ala701Val) (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

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Val16

Predicted Molecular Mass

138 kDa

SDS-PAGE

145-168 kDa under reducing conditions.

Activity

Measured by flow cytometry for its ability to bind HEK293 human embryonic kidney cells transfected with human ACE-2 at 0.25-1.00 µg/mL (100 µL/well).
Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Images for Recombinant SARS-CoV-2 S GCN4-IZ Alexa Fluor® 488 Protein

Detection of ACE-2 on HEK293 Transfectants with Recombinant SARS-CoV-2 B.1.351 Spike (GCN4-IZ) His-tag Alexa Fluor® 488 by Flow Cytometry.

HEK293 human embryonic kidney cells transfected with human ACE-2 were stained with (A) 1 µg/mL (100 µL/well) Recombinant SARS-CoV-2 B.1.351 Spike (GCN4-IZ) His-tag Alexa Fluor® 488 (Catalog # AFG10786) or (B) unstained.

Recombinant SARS-CoV-2 B.1.351 Spike (GCN4-IZ) His-tag Alexa Fluor® 488 Protein SDS-PAGE.

2 μg/lane of Recombinant SARS-CoV-2 B.1.351 Spike (GCN4-IZ) His-tag Alexa Fluor® 488 Protein (Catalog # AFG10786) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 145-168 kDa.

Formulation, Preparation, and Storage

AFG10786
Formulation Supplied as a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

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 (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 (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 ACE-2 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 amino acid substitutions N501Y, K417N, and E484K in the RBD raised the most concerns. This B.1.351 lineage, also known and 501Y.V2 variant, was first identified in the Eastern Cape province of South Africa in December 2020 and spread quickly to become the most dominant strain in the second COVID wave in South Africa (11). Two of these mutations K417N and E484K locate at the receptor binding motif (RBM) and are not found in other variants (11). The N501Y mutation is also found in London (B.1.1.7 lineage) and Brazil (P.1 lineage). The B.1.351 lineage is reported to enter cells more easily due to its enhanced affinity to ACE-2 receptor (12). It is reported to reduce the efficacy of neutralizing antibody (12, 13).

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://doi.org/10.1101/2020.03.14.988345.
  11. Tegally, H. et al. (2020) bioRxiv. Doi: https://doi.org/10.1101/2020.12.21.20248640.
  12. Nelson, G. et al. (2021) bioRxiv. https://doi: 10.1101/2021.01.13.426558.
  13. Wibmer, C.K. et al. (2021) bioRxiv. https://doi: 10.1101/2021.01.18.427166.

Long Name

Spike Protein

Alternate Names

S Protein

Entrez Gene IDs

918758 (HCoV-229E); 2943499 (HCoV-NL63); 39105218 (HCoV-OC43); 37616432 (MERS-CoV); 1489668 (SARS-CoV); 43740568 (SARS-CoV-2)

Gene Symbol

S

UniProt

YP_009724390.1

Additional Spike Products

Product Documents for Recombinant SARS-CoV-2 S GCN4-IZ Alexa Fluor® 488 Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant SARS-CoV-2 S GCN4-IZ Alexa Fluor® 488 Protein

This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.
This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

For research use only

Related Research Areas

Infectious Diseases

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