SARS-CoV-2 Spike Antibody - BSA Free

Western Blot: SARS-CoV-2 Spike Antibody [NBP2-41058] - Validation in Spike Transfected 293 Cells Loading: 15 ug per lane of 293 cell lysate. SARS-CoV-2 Spike(1ug/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

Immunohistochemistry-Paraffin: SARS-CoV-2 Spike Antibody [NBP2-41058] - Immunohistochemistry Validation of SARS-CoV-2 Spike in COVID-19 Patient Lung. Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti-SARS-CoV-2 Spike S2 antibody (NBP2-41058, 0.5 ug/mL). Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Strong spike protein signal was observed in macrophages of COVID-19 patient lung.

Immunohistochemistry-Paraffin: SARS-CoV-2 Spike Antibody [NBP2-41058] - Immunohistochemistry Validation of SARS-CoV-2 Spike in COVID-19 Patient Lung. Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti-SARS-CoV-2 Spike S2 antibody (NBP2-41058, 0.5 ug/mL). Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Strong spike protein signal was observed in macrophages and airway epithelium of COVID-19 patient lung, but not in non-COVID-19 patient lung.

ELISA: SARS-CoV-2 Spike Antibody - BSA Free [NBP2-41058] - Figure 14 ELISA TestAntibodies: SARS-CoV-2 (COVID-19) Spike antibody, (1 u/mL). A direct ELISA was performed using immunogen or control peptide as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 0.5 ng/mL to 1000 ng/mL.

Immunocytochemistry/ Immunofluorescence: SARS-CoV-2 Spike Antibody - BSA Free [NBP2-41058] - Validation of SARS-CoV-2 (COVID-19) Spike in 293T Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed 293T cells labeling SARS-CoV-2 (COVID-19) Spike with at 1 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

• Reactivity: V, V
• Applications: WB, ELISA, ICC/IF, IHC
• 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 20 amino acids near the carboxy terminus of SARS-CoV-2 Spike glycoprotein. The immunogen is located within the last 50 amino acids of SARS-CoV-2 Spike protein.
• Predicted Species: SARS-CoV (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: S
• Purity: Peptide affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1 ug/mL
  • Immunocytochemistry/ Immunofluorescence 1 ug/ml
  • Immunohistochemistry 0.2 ug/mL
  • Immunohistochemistry-Paraffin
  • Western Blot 1 ug/ml
• Application Notes: The immunogen for this is within the last 50 aa of the spike protein - a peptide corresponding to 20 amino acids near the carboxy terminus of SARS-CoV-2 (COVID-19) Spike glycoprotein. The Extracellular domain (ECD) is from aa 1 to 1208 (full length 1273aa). Therefore, this antibody detects the transmembrane and cytoplasm domains at the C terminus, but does not detect the ECD (which is the region expressed in many commercially available spike proteins). NBP2-41058 can be used for the detection of full length spike protein and spike protein in COVID-19 patient samples.

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 (NBP2-41058)(1)

Publication using NBP2-41058

• Kiyan Y, Schultalbers A, Chernobrivaia E et al. Calcium dobesilate reduces SARS-CoV-2 entry into endothelial cells by inhibiting virus binding to heparan sulfate Scientific reports 2022-10-07 [PMID: 36207386] (ICC/IF, IHC-Fr)

Bioinformatics

• Gene Symbol: S
• Uniprot:
  • QHD43416()