Anti-SARS-CoV-2 Spike RBD Antibody, clone 8B12-C2

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. (Ref.: Abe, KT., et al. (2020). JCI Insight. 5(19); e142362).

Clone 8B12-C2 is a mouse monoclonal antibody that detects spike glycoprotein of SARS-CoV-2. It targets an epitope within a highly conserved region in the receptor binding domain.

A KLH-conjugated linear peptide from a highly conserved sequence localized within the receptor binding domain of SARS-CoV-2.

Quality Control Testing

Evaluated by Western Blotting in lysate from HEK293T cells expressing full-length spike protein of SARS-CoV-2.

Western Blotting Analysis: A 1:500 dilution of this antibody detected spike glycoprotein in lysate from HEK293T expressing full-length SARS-CoV-2 spike protein, but not in lysate from wild type HEK393T cells.

Tested Applications

Western Blotting Analysis: A 1:400 dilution from a representative lot detected SARS-CoV-2 Spike protein in whole cell lysates from HEK293T cells expressing full-length spike protein of SARS-CoV-2, but not in untransfected cells. (Courtesy of Stefan Schüchner, Ingrid Mudrak, Ingrid Frohner, and E. Ogris (Max Perutz Labs, Medical University of Vienna, Austria).

ELISA Analysis: Various dilutions of a hybridoma culture supernatant for clone 8B12-C2 detected native SARS-CoV-2 spike protein. ( (Courtesy of Stefan Schüchner, Ingrid Mudrak, Ingrid Frohner, and E. Ogris (Max Perutz Labs, Medical University of Vienna, Austria).

Immunoprecipitation Analysis: A representative lot immunoprecipitated spike glycoprotein of SARS-CoV-2. (Courtesy of Stefan Schüchner, Ingrid Mudrak, Ingrid Frohner, and E. 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

Anti-SARS-CoV-2 Spike RBD, clone 8B12-C2, Cat. No. MABF2783, is a mouse monoclonal antibody that detects an epitope of spike protein conserved in the α, β, γ, δ, and ο variants of SARS-CoV-2 and is used in ELISA, Immunoprecipitation & Western Blotting.

Purified mouse monoclonal antibody IgG1 in PBS without azide.

Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

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

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.