Recombinant SARS-CoV-2 A.23.1 Spike GCN4-IZ His Protein, CF

First Detected in Uganda. +E484K
Catalog # Availability Size / Price Qty
10860-CV-100
Recombinant SARS-CoV-2 A.23.1 Spike (GCN4-IZ) His-tag Protein Binding Activity.
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Recombinant SARS-CoV-2 A.23.1 Spike GCN4-IZ His Protein, CF Summary

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
SARS-CoV-2 Spike
(Val16-Lys1211)(Arg102Ile, Phe157Leu, Val367Phe, Glu484Lys, Gln613His, Pro681Arg)(Arg682Ser, Arg685Ser, Lys986Pro, Val987Pro)
Accession # YP_009724390.1
GCN4-IZ6-His tag
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Val16
Predicted Molecular Mass
138 kDa
SDS-PAGE
140-166 kDa, under reducing conditions.

Product Datasheets

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10860-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.

10860-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

Binding Activity View Larger

Recombinant SARS-CoV-2 A.23.1 Spike (GCN4-IZ) His-tag (Catalog # 10860-CV) binds Recombinant Human ACE-2 His-tag (933-ZN) in a functional ELISA.

SDS-PAGE View Larger

2 μg/lane of Recombinant SARS-CoV-2 A.23.1 Spike (GNC4-IZ) His-tag Protein (Catalog # 10860-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 140-166 kDa.

Surface Plasmon Resonance (SPR) Surface plasmon resonance (SPR) sensorgram of Human ACE-2 binding to SARS-CoV-2 mutant Spike protein A.23.1 variant E484K View Larger

Recombinant SARS-CoV-2 A.23.1 variant with E484K Spike protein His-tag (Catalog #10860-CV) 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=2.29 nM. (Biacore T200).

Reconstitution Calculator

Reconstitution Calculator

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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 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). A metallopeptidase, angiotensin-converting enzyme 2 (ACE-2), has been identified as a functional receptor for SARS-CoV-2 through interaction with a receptor binding domain (RBD) located at the C-terminus of S1 subunit (6, 7). 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. A SARS-CoV-2 variant (A.23.1) carrying the amino acid substitution F157L, V367F, E484K, Q613H and P681R in the spike protein region was identified as a prevalent strain in Uganda and is now spread to 26 counties (8). Whether these mutations in spike protein would cause more severe symptom or decrease the efficacy of vaccine-induced immunity is still under investigation.

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. Li, W. et al. (2003) Nature 426:450.
  7. Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
  8. Bugembe, D.L. et al. (2021) medRxiv. DOI: 10.1101/2021.02.08.21251393.
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

FAQs

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