Recombinant SARS-CoV-2 G476S Spike RBD His-tag Protein, CF

Catalog #: 10627-CV Datasheet / COA / SDS

Gly476Ser Point Mutation

Catalog #	Availability	Size / Price	Qty
10627-CV-100

Recombinant SARS-CoV-2 G476S 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 G476S 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 (Gly476Ser), with a C-terminal 6-His tag.

Accession #

YP_009724390.1

N-terminal Sequence
Analysis

Arg319

Predicted Molecular Mass

26 kDa

SDS-PAGE

32-40 kDa, under reducing and non-reducing conditions

Product Datasheets

10627-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.

10627-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 G476S Spike RBD His-tag (Catalog # 10627-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 G476S Spike RBD His-tag (Catalog # 10627-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 32-40 kDa.

Surface Plasmon Resonance (SPR)

View Larger

Recombinant SARS-CoV-2 Spike RBD G476S 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 0.18 nM and 94.3 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=2.432 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. A SARS-CoV-2 variant carrying the aa substitution G476S mutation is one of the more prevalent mutations found in the RBD (8, 9). While the G476S mutation is directly in the ACE2 binding region, there does not appear to be an effect on binding affinity (8, 9).

References