SARS-CoV-2 Spike Antibody - BSA Free

ELISA: SARS-CoV-2 Spike Antibody [NBP3-11940] - Validation of SARS-Cov-2 Spike Antibody with Spike S1 Protein of SARS-CoV-2 Variants. Coating Antigen: SARS-CoV-2 spike S1 proteins WT, alpha variant (B.1.1.7), beta variant (B.1.351), gamma variant (P.1), delta variant (B.1.617.2), and omicron variant (B.1.1.529), 1 ug/mL, incubated at 4C overnight. Detection Antibody: SARS-CoV-2 Spike antibody NBP3-11940, 0.2-1 ug/mL incubated at RT for 1 hr. Secondary Antibody: Goat anti-rabbit HRP at 1:20,000, incubate at RT for 1 hr. NBP3-11940 can detect spike S1 protein for all variants except omicron.

Western Blot: SARS-CoV-2 Spike Antibody [NBP3-11940] - Western Blot Validation with SARS-CoV-2 (COVID-19) Spike S1 Recombinant Protein. Loading: 50 ng per lane of SARS-CoV-2 (COVID-19) Spike S1 recombinant protein. Antibodies: SARS-CoV-2 Spike, NBP3-11940, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 ug/mL and Lane 2: 2 ug/mL.

Immunocytochemistry/Immunofluorescence: SARS-CoV-2 Spike Antibody [NBP3-11940] - Immunofluorescence Validation of SARS-CoV-2 (COVID-19) Spike (cleavage site) in 293 Transfected Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed Spike transfected 293cells labeling SARS-CoV-2 (COVID-19) Spike (cleavage site) with NBP3-11940 at 10 mg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Western Blot: SARS-CoV-2 Spike Antibody [NBP3-11940] - Overexpression Validation in Spike Transfected 293 Cells. Loading: 10 ug per lane of 293 cell lysate. Antibodies: SARS-CoV-2 Spike, NBP3-11940 (0.5 ug/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: WT 293 cells and Lane 2: SARS-CoV-2 Spike overexpressed 293 cells

Western Blot: SARS-CoV-2 Spike Antibody [NBP3-11940] - Western Blot Validation with SARS-CoV-2 (COVID-19) Spike Recombinant Protein. Loading: 30 ng per lane of SARS-CoV-2 (COVID-19) Spike RBD+SD1+SD2 recombinant protein. Antibodies: SARS-CoV-2 Spike, NBP3-11940, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 ug/mL, Lane 2: 2 ug/mL and Lane 3: 4 ug/mL.

ELISA: SARS-CoV-2 Spike Antibody [NBP3-11940] - Detection of SARS-CoV-2 UK Variant Spike S1 Protein with Spike S1 Antibodies by Direct ELISA. Coating Antigen: SARS-CoV-2 spike S1 proteins, including WT and UK variant (B.1.1.7), 1 ug/mL Incubate at 4C overnight. Detection Antibodies: SARS-CoV-2 Spike Antibody, NBP3-11940, dilution: 0.1-1000 ng/mL, incubate at RT for 1 hr. Secondary Antibodies: Goat anti-rabbit HRP at 1:20,000, incubate at RT for 1 hr. Immunogen region of antibody NBP3-11940 includes site 681P that was mutated in B.1.1.7.. SARS-CoV-2 Spike antibody (NBP3-11940) has lower binding affinity for variant S1 protein as compared to WT S1 protein.

ELISA: SARS-CoV-2 Spike Antibody [NBP3-11940] - ELISA Validation of SARS-CoV-2 Spike Antibody Pair. A sandwich ELISA was performed using the anti-SARS-CoV-2 Spike antibodies NBP3-11940 (10 ug/mL ) as the capture antibody. Biotin-labled anti-SARS-COV-2 Spike S1 antibodies (1 ug/mL ) and streptavidin-HRP (0.1 ug/mL ) were used for detection. Detection range is from 0.8 ng to 200 ng (SARS-CoV-2 Spike S1 Protein).

ELISA: SARS-CoV-2 Spike Antibody [NBP3-11940] - ELISA Validation with SARS-CoV-2 (COVID-19) Spike Recombinant Protein. Antibodies: SARS-CoV-2 Spike, NBP3-11940. A direct ELISA was performed using SARS-CoV-2 (COVID-19) Spike RBD+SD1+SD2 recombinant protein, as coating antigen and the anti-SARS-CoV-2 Spike as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 8 ng/mL to 1000ng/mL.

ELISA: SARS-CoV-2 Spike Antibody [NBP3-11940] - ELISA Validation with SARS-CoV-2 (COVID-19) Spike S1 Recombinant Protein. Antibodies: SARS-CoV-2 Spike antibody, NBP3-11940 (1 ug/mL). A direct ELISA was performed using SARS-CoV-2 (COVID-19) Spike S1 recombinant protein as coating antigen and the anti-SARS-CoV-2 Spike antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 0.3 ng/mL to 1000ng/mL.

• Reactivity: V
• Applications: WB, ELISA, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

SARS-CoV-2 Spike Antibody - BSA Free Summary

• Immunogen: Antibody was raised against a peptide corresponding to 12 amino acids near the center of SARS-CoV-2 Spike glycoprotein. The immunogen is located within 650-700 amino acids of SARS-CoV-2 Spike protein.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: S
• Purity: Peptide affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Immunocytochemistry/ Immunofluorescence 20 ug/mL
  • Sandwich ELISA 10 ug/ml
  • Western Blot 0.5-4 ug/mL

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Peptide affinity purified

Alternate Names for SARS-CoV-2 Spike Antibody - BSA Free

• 2019-nCoV S Protein
• 2019-nCoV Spike
• COVID-19 Spike
• E2
• Human coronavirus spike glycoprotein
• Peplomer protein
• S glycoprotein
• S Protein
• SARS-COV-2 S protein
• SARS-COV-2 Spike glycoprotein
• SARSCOV2 Spike protein
• SARS-CoV-2
• Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein
• Spike glycoprotein
• Spike
• surface glycoprotein

Background

The SARS-CoV-2 Spike protein is one of the four major structural proteins of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causative agent of COVID-19 (1,2). The spike protein is the largest of the structural proteins, which also include the membrane (M), envelope (E), and nucleocapsid (N) proteins (1,2). The SARS-CoV-2 spike protein is a 1273 amino acid (aa) heterotrimeric class I fusion protein with each monomer having a theoretical molecular weight of approximately 180 kDa (1). The club-shaped spike protein contains several functional regions and domains including the S1 globular head region which contains the N-terminal receptor-binding domain (RBD) and the S2 stem region that contains the C-terminal fusion domain, two heptad regions, a transmembrane domain, and a cytoplasmic tail (1,2). The viral spike protein is critical for attachment of the virus with the host cell, resulting in fusion and virus entry into the cell (1,2). More specifically, the RBD of the spike protein is responsible for binding to the cell surface receptor angiotensin converting enzyme 2 (ACE2) (1,2). This spike-ACE2 interaction results in a conformational change permitting furin cleavage between the S1 and S2 domains and then cleavage at S2' by TMPRRS2, or another protease, allowing membrane fusion (1,2).

Given the critical role of the spike protein RBD in the interaction with the ACE2 receptor and viral entry, a number of neutralizing antibodies against the RBD have been developed as potential therapeutics for treating COVID-19 (3). These antibodies bind the RBD domain on the S1 subunit inhibiting the interaction with ACE2 (3). However, more studies need to be done as neutralizing antibodies can result in antibody-dependent enhancement, in which the viral entry and replication within the host cell is increased (4). One potential way to combat antibody-dependent enhancement is the use of nanobodies (4). Furthermore, there are currently several vaccine strategies that are in clinical trials, or recently federally approved, that utilize the spike protein in different forms (e.g. full length, S1 RBD, RBD-Fc, N-terminal) for protecting against SARS-CoV-2 infection (4,5). These vaccine strategies include DNA vaccines, viral vector-based vaccines, RNA vaccines, and subunit vaccines (4,5).

References

1. Pillay T. S. (2020). Gene of the month: the 2019-nCoV/SARS-CoV-2 novel coronavirus spike protein. Journal of Clinical Pathology. https://doi.org/10.1136/jclinpath-2020-206658

2. Malik Y. A. (2020). Properties of Coronavirus and SARS-CoV-2. The Malaysian Journal of Pathology.

3. Ho M. (2020). Perspectives on the development of neutralizing antibodies against SARS-CoV-2. Antibody Therapeutics. https://doi.org/10.1093/abt/tbaa009

4. Samrat, S. K., Tharappel, A. M., Li, Z., & Li, H. (2020). Prospect of SARS-CoV-2 spike protein: Potential role in vaccine and therapeutic development. Virus Research. https://doi.org/10.1016/j.virusres.2020.198141

5. Sternberg, A., & Naujokat, C. (2020). Structural features of coronavirus SARS-CoV-2 spike protein: Targets for vaccination. Life Sciences. https://doi.org/10.1016/j.lfs.2020.118056

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for SARS-CoV-2 Spike Antibody (NBP3-11940)(1)

Publication using NBP3-11940

• V Bezzerri, V Gentili, M Api, A Finotti, C Papi, A Tamanini, C Boni, E Baldisseri, D Olioso, M Duca, E Tedesco, S Leo, M Borgatti, S Volpi, P Pinton, G Cabrini, R Gambari, F Blasi, G Lippi, A Rimessi, R Rizzo, M Cipolli SARS-CoV-2 viral entry and replication is impaired in Cystic Fibrosis airways due to ACE2 downregulation Nature Communications, 2023-01-10;14(1):132. 2023-01-10 [PMID: 36627352]

Bioinformatics

• Gene Symbol: S