ANTI-MERS-COV NUCLEOPROTEIN. CLONE 46 (C15-1886-688)

Specificity

Clone 6D4-E2 is a mouse monoclonal antibody that detects the alpha variant of SARS-CoV-2 with N501Y mutation in Spike protein.

Immunogen

A recombinant fusion protein with a scaffold containing a short spike peptide GFQPTYGVGYQ.

Application

Quality Control Testing

Evaluated by Western Blotting in lysate from HEK293T cells transiently transfected with SARS-CoV-2 spike B.1.351, beta.

Western Blotting Analysis: A 1:500 dilution of this antibody detected SARS-CoV-2 spike protein with N501Y mutation in lysate from HEK293T cells transiently transfected with SARS-CoV-2 spike B.1.351, Beta, but not in wild-type HEK293T cells.

Tested Applications

Western Blotting Analysis: A representative lot detected SARS-CoV-2 spike N501Y in HEK293T transiently transfected with various SARS-CoV-2 variants (Data courtesy of Stefan Schüchner and Egon Ogris (Max Perutz Labs, Medical University of Vienna, Austria).

Immunoprecipitation Analysis: A representative lot detected SARS-CoV-2 spike N501Y in Whole cell lysates of HEK293T cells (untransfected or transiently transfected with SARS-CoV-2 spike WT or B.1.351 variant) (Data courtesy of Stefan Schüchner and Egon Ogris (Max Perutz Labs, Medical University of Vienna, Austria).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Target description

Spike glycoprotein (UniProt: P0DTC2; also known as S glycoprotein, E2, Peplomer protein) is encoded by the S (also known as 2) gene (Gene ID: 43740568) in Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The SARS-CoV-2 is a positive-strand RNA virus that causes severe respiratory syndrome in human. The mature SARS-CoV-2 contains 4 structural proteins: Envelope (E), Membrane (M), Nucleocapsid (N), and the Spike protein (S). E and M proteins help in viral assembly and N protein is needed for RNA synthesis. The S protein is a single-pass type I, homotrimeric, membrane glycoprotein that is responsible for virus binding and entry into host cell. It is synthesized with a signal peptide (aa 1-12), which is subsequently cleaved off to generate the mature protein that contains an extracellular domain (aa 13-1213), a transmembrane domain (aa 1214-1234), and a cytoplasmic domain 1235-1273). The S protein is further processed into S1 (aa 13-685) and S2 (aa 686-1273) subunits by host cell furin. The presence of a furin polybasic cleavage site in S protein from SARS-CoV-2 sets it apart from S protein in SARS-CoV that possesses a monobasic S1/S2 cleavage site. The S1 subunit has the receptor binding domain (RBD; aa 319-541) that mediates entry of SARS-CoV-2 into sensitive cells through the peptidase domain of host Angiotensin-converting enzyme 2 (ACE2) with high affinity (KD = 15 nM). The S2 domain (aa 686-1213), which is reported to be well conserved, is responsible for membrane fusion. Over the course of the pandemic, SARS-CoV2 has consistently mutated giving rise to various mutants. The Alpha variant of SARS-CoV-2 contains 19 nonsynonymous mutations across its viral genome, including 8 substitutions or deletions in the spike protein. The N501Y mutation in Alpha variant results in enhanced viral replication in human airway epithelial cells and is a critical determinant of enhanced infection of the upper airway and transmission. This mutation resides in the ACE2 interface of the receptor binding domain (RBD). Clone 6D4-E2 specifically detects the SARS-CoV-2 Spike mutation N501Y. (Ref.: Liu, Y., et al. (2022). Nature. 602(7896); 294-299; Cheng, L., et al. (2021). Virol. J. 18; 87).

Physical form

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Reconstitution

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.