Recombinant HCoV-NL63 Spike RBD His-tag Protein, CF Summary
Ala475-Asp634, with a C-terminal 6-His tag
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.
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.
|Formulation||Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.|
|Reconstitution||Reconstitute at 200 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.
Recombinant HCoV-NL63 Spike RBD His-tag Protein (Catalog # 10605-CV) binds Recombinant Human ACE-2 Fc Chimera Protein (10544-ZN) in a functional ELISA.
2 μg/lane of Recombinant HCoV-NL63 Spike RBD His-tag (Catalog # 10605-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 30-38 kDa.
Background: Spike RBD
HCoV-NL63, a virus first isolated from a child suffering from respiratory disease in 2003, belongs to a family of viruses known as coronaviruses 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) (1, 2). Other well-known human coronaviruses include three viruses that cause relatively mild respiratory disease: HCoV-229E, HCoV-HKU1 and HCov-OC43, plus three viruses that cause the Severe Acute Respiratory Syndrome (SARS-CoV), the Middle East Respirator Syndrome (MERS-CoV), and the global pandemic Covid-19 (SARS-CoV2). HCov-NL63 Spike Protein (S Protein) is a glycoprotein that mediates membrane fusion and viral entry. As with most coronaviruses, proteolytic cleavage of the S protein generates two distinct peptides, S1 and S2 subunits. The S1 subunit is focused on attachment of the protein to the host receptor while the S2 subunit is involved with cell fusion. Although HCoV-NL63 S protein shares high homology (56%) with HCoV-229E, it does not employ CD13 (aminopeptidase N) as the receptor like HCoV-229E. Instead, HCoV-NL63 engages Angiotensin-Converting Enzyme 2 (ACE-2), the same receptor as SARS-CoV and SARS-CoV2, for cellular entry and replication (3). The receptor binding domain (RBD) of HCoV-NL63 is located at C-terminal region of S1 subunit (4, 5). Although NL63-CoV and SARS-CoV do not share structural homology in RBD region, they bind an overlapping region of ACE-2 (6, 7).
- Van der Hoek, L. et al. (2004) Nat. Med. 10:368.
- Fouchier, R.M. et al. (2004) Proc. Natl. Acad. Sci. U.S.A. 101:6212.
- Hofmann, H. et al. (2005) Proc. Natl. Acad. Sci. U.S.A. 102:7988.
- Hofmann, H. et al. (2006) J. Virol. 80:8639.
- Lin, H. et al. (2008) J. Gen. Virol. 89:1015.
- Li, W. et al. (2007) Virology 367:367.
- Wu, K. et al. (2009) Proc. Natl. Acad. Sci. U.S.A. 106:19970.
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