Recombinant SARS-CoV-2 F490S Spike RBD His-tag Protein, CF

Catalog #: 10815-CV Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
10815-CV-100

Recombinant SARS-CoV-2 F490S Spike RBD His-tag Protein Binding Activity.

3 Images

Product Details

Reviews

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant SARS-CoV-2 F490S Spike RBD His-tag 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 RBD protein
Arg319-Phe541 (Phe490Ser), with a C-terminal 6-His tag

Accession #

YP_009724390.1

N-terminal Sequence
Analysis

Arg319

Predicted Molecular Mass

26 kDa

SDS-PAGE

26-38 kDa, under reducing conditions.

Product Datasheets

10815-CV

Product Datasheet

COA

SDS

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.

10815-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 F490S Spike RBD His-tag (Catalog # 10815-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 F490S Spike RBD His-tag (Catalog # 10815-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 26-38 kDa.

Surface Plasmon Resonance (SPR)

View Larger

Recombinant SARS-CoV-2 Spike RBD protein F490S His-tag 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= 1.52 nM.

Reconstitution Calculator

Background: Spike RBD

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 (ACE2), 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 RBD of SARS-CoV-2 shares 73% aa identity with the RBD of the SARS-CoV-1, but only 22% amino acid (aa) identity with the RBD of MERS. SARS-CoV-2 variants carrying the aa substitution F490S in the RBD was reported to escape neutralization of antibodies (8, 9).

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.
Li, W. et al. (2003) Nature 426:450.
Wong, S.K. et al. (2004) J. Biol. Chem. 279:3197.
Weisblum, Y. et al. (2020) bioRxiv. https://doi.org/10.1101/2020.07.21.214759.
Liu, Z. et al. (2021) Cell Host & Mircobe 29:477.