Biotinylated Recombinant SARS-CoV-2 B.1.1.7 Spike Protein S1 (carrier-free)

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Regulatory Status
RUO
Other Names
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 protein, UK variant, S1, Coronavirus spike protein
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SARS-CoV-2-B117-_Spike-Protein-S1-_RECOM_BA_1_042123
When recombinant human ACE2-Fc (Cat. No. 793202) is immobilized at 1 µg/mL, biotinylated recombinant SARS-CoV-2 B.1.1.7 Spike protein S1 binds in a dose-dependent manner. The ED50 range for this effect is 3 - 18 ng/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
  • SARS-CoV-2-B117-_Spike-Protein-S1-_RECOM_BA_1_042123
    When recombinant human ACE2-Fc (Cat. No. 793202) is immobilized at 1 µg/mL, biotinylated recombinant SARS-CoV-2 B.1.1.7 Spike protein S1 binds in a dose-dependent manner. The ED50 range for this effect is 3 - 18 ng/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
  • SARS-CoV-2-B117-_Spike-Protein-S1-_RECOM_BA_2_042123
    Stability testing for Biotinylated recombinant SARS-CoV-2 B.1.1.7 Spike protein S1. Biotinylated recombinant SARS-CoV-2 B.1.1.7 Spike protein S1 was aliquoted in 10 mM NaH2PO4, pH 7.5 at 0.2 mg/mL and one aliquot was kept at 4°C (Control), and another was frozen and thawed four times (4x Freeze/Thaw). The samples were tested by immobilizing recombinant human ACE2- Fc (Cat. No. 793202) at 1 µg/mL to which biotinylated recombinant SARS-CoV-2 B.1.1.7 Spike protein S1 then binds in a dose-dependent manner. The ED50 range for this effect is 3 - 18 ng/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
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560304 25 µg 344€
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560306 100 µg 740€
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Description

The severe acute respiratory syndrome coronavirus (SARS-CoV-2) is a respiratory virus which causes coronavirus disease 2019 (COVID-19). The coronavirus spike (S) glycoprotein is a class I viral fusion protein on the outer envelope of the virion that plays a critical role in viral infection by recognizing host cell receptors and mediating fusion of the viral and cellular membranes. The S glycoprotein is synthesized as a precursor protein consisting of ~1,300 amino acids that is then cleaved into an amino (N)-terminal S1 subunit (~700 amino acids) and a carboxyl (C)-terminal S2 subunit (~600 amino acids). Three S1/S2 heterodimers assemble to form a trimer spike protruding from the viral envelope. The S1 subunit contains a receptor binding domain (RBD) that can specifically bind to angiotensin-converting enzyme 2 (ACE2), the receptor on target cells. Triggered by receptor binding, proteolytic processing and/or acidic pH in the cellular compartments, the class I viral fusion protein undergoes a transition from a metastable pre-fusion state to a stable post-fusion state during infection, in which the receptor-binding subunit is cleaved, and the fusion subunit undergoes large-scale conformational rearrangements to expose the hydrophobic fusion peptide, induce the formation of a six-helix bundle, and bring the viral and cellular membranes close for fusion. The trimeric SARS coronavirus (SARS-CoV-2) S glycoprotein consisting of three S1-S2 heterodimers binds the cellular receptor angiotensin-converting enzyme 2 (ACE2) and mediates fusion of the viral and cellular membranes through a pre- to post-fusion conformation transition. The emergence of the B.1.1.7 variant of SARS-CoV-2, also known as the variant of concern (VOC) 202012/01 strain, was first identified in the United Kingdom (UK) in 2020. Additionally, the B.1.1.7 was significantly more transmissible than other preexisting variants. The B.1.1.7 variant contains a significant mutation of interest in the RBD, the asparagine (N) to tyrosine (Y) mutation at position 501 (N501Y), which has been shown to result in enhanced binding affinity for hACE-2.

Product Details
Technical Data Sheet (pdf)

Product Details

Source
Biotinylated recombinant SARS-CoV-2 B.1.1.7 Spike protein S1, amino acid Val16-Arg685 (Accession No. QHD43416.1), with (HV69-70del, Y144del, N501Y, A570D, P681H) and a C-terminal 8His-Avi tag was expressed in CHO cells. SARS-CoV-2 B.1.1.7 Spike Protein S1 Avi-tag was site-specifically biotinylated by enzyme BirA.
Molecular Mass
The 703 amino acid recombinant protein has a predicted molecular mass of approximately 78 kD. The DTT-reduced and non-reduced protein migrates at approximately 80 - 115 kD by SDS-PAGE. The predicted N-terminal amino acid is Val.
Purity
> 95%, as determined by Coomassie stained SDS-PAGE.
Formulation
0.22 µm filtered protein solution is in 10mm NaH2PO4, pH 7.5
Endotoxin Level
Less than 0.1 EU per µg cytokine as determined by the LAL method.
Concentration
25 µg size is bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & Handling
Unopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
Activity
When recombinant human ACE2- Fc (Cat. No. 793202) is immobilized at 1 µg/mL, biotinylated recombinant SARS-CoV-2 B.1.1.7 Spike protein S1 binds in a dose-dependent manner. The ED50 range for this effect is 3 - 18 ng/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
Application

Bioassay

Application Notes

BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at tech@biolegend.com

Antigen Details

Structure
Monomer
Distribution

SARS-CoV-2

Function
Attaches the virion to the cell membrane by interacting with host receptor (ACE2) initiating the infection
Interaction
Ciliated cells in nasal mucosa and bronchus
Ligand/Receptor
ACE2
Bioactivity
Measured by its ability to bind recombinant human ACE2
Cell Type
B cells, Endothelial cells, Macrophages, Monocytes, NK cells, T cells, Th1
Biology Area
Adaptive Immunity, Immunology, Innate Immunity
Antigen References
  1. Lu R, et al. 2020. Lancet. 395:565-574.
  2. Li F. 2016. Annu Rev Virol. 29;3(1):237-261.
  3. Belouzard S, et al. 2012. Viruses. 4:1011-33.
  4. Song W, et al. 2018. PLoS Pathog. 14:e1007236
  5. Li F, et al. 2020. Nature. 581:221.
  6. TJ Yang, et al. 2021. Nature structural & Molecular Biology. 28:731-739.
  7. Kidd M, et al. 2021. The Journal of Infectious Diseases. 224:928-929.
  8. Davis NG, et al. 2021. Science. 372 (6538): eabg305.
Gene ID
43740568 View all products for this Gene ID
UniProt
View information about S1 on UniProt.org
Go To Top Version: 1    Revision Date: 04-21-2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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