Recombinant MERS-CoV Spike RBD His-tag Protein, CF

Catalog #: 10621-CV Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
10621-CV-100

Recombinant MERS-CoV Spike RBD His-tag Protein Binding Activity.

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Product Details

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

Recombinant MERS-CoV 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 DPPIV/CD26 (High Purity Dimer) (Catalog # 9168-SE).

Source

Chinese Hamster Ovary cell line, CHO-derived mers-cov Spike RBD protein

MERS-CoV Spike RBD (Glu367-Tyr606) Accession # YP_007188579.1	HP	GGGSGGGSGGGS	HHHHHH
N-terminus			C-terminus

Accession #

YP_007188579.1

N-terminal Sequence
Analysis

Glu367 & Lys369

Predicted Molecular Mass

28 kDa

SDS-PAGE

35-45 kDa, under reducing conditions

Product Datasheets

10621-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.

10621-CV

Formulation	Supplied as a 0.2 μm filtered solution in PBS with Trehalose.
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	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.

Scientific Data

Binding Activity

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Recombinant MERS-CoV Spike RBD His-tag (Catalog # 10621-CV) binds Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein (9168-SE) in a functional ELISA.

SDS-PAGE

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2 μg/lane of Recombinant MERS-CoV Spike RBD His-tag Protein (Catalog # 10621-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 35-45 kDa.

Flow Cytometry

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In a functional flow cytometry test, (A) Recombinant MERS-CoV Spike RBD His-tag Protein (Catalog # 10621-CV) binds to HEK293 human embryonic kidney cell line transfected with recombinant human CD26/DPPIV and EGFP. Ligand binding was detected by staining cells with APC-conjugated anti-His Monoclonal Antibody (IC050A), which does not stain the cells in the absence of recombinant protein (B).

Reconstitution Calculator

Background: Spike RBD

MERS-CoV (also known as HCoV-EMC), which causes the Middle East Respiratory Syndrome (MERS), was first reported in Saudi Arabia in 2012 as a novel coronavirus (1). Coronaviruses are a family of viruses that are commonly comprised of a large plus-strand RNA genome and four structural proteins: Spike protein (S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N). There are two well-known human coronavirus families that infect humans: Alpha coronaviruses which includes HCoV-229E and HCoV-NL63; beta coronaviruses that includes MERS-CoV, HCov-OC43, Severe Acute Respiratory Syndrome (SARS-CoV), and global pandemic Covid-19 (SARS-CoV2) (2). The MERS-CoV Spike Protein (S Protein) is a glycoprotein that mediates membrane fusion and viral entry, and it consists of two subunits, S1 and S2. The S1 subunit is focused on attachment of the protein to the host receptor while the S2 subunit is involved with cell fusion (3). Located within the S1 subunit is the receptor binding domain (RBD). The RBD is responsible for the binding of MERS-CoV to dipeptidyl peptidase IV (DPP4, also known as human CD26) (4). The RBD of MERS-CoV shares 24% and 21% amino acid sequence (aa) identity with SARS-CoV RBD and SARS-Cov2 RBD, respectively. The low aa sequence identity is consistent with the finding that MERS-CoV and SARS-CoV bind different cellular receptors (4). The S1 subunit, especially the RBD region, of MERS-CoV was commonly targeted for vaccinations or antiviral therapies (5-7).

References

Zaki, A.M. et al. (2012) N. Engl. J. Med. 367:1814.
Ogimi, C. et al. (2020) J Pediatric Infect Dis Soc doi: 10.1093/jpids/piaa037.
Li, Y. et al. (2019) Engineering. 5:940.
Raj, V.S. et al. (2013) Nature 495:251.
Corti, D. et al. (2016) J. Infect. Public Health 9:231.
Tang, X.C. et al. (2014) Proc. Natl. Acad. Sci. USA 111:E2018.
Jiang, L. et al. (2014) Sci. Transl. Med. 6:234ra59.