Flavivirus group antigen Antibody (D1-4G2-4-15 (4G2)) - Chimeric - Azide and BSA Free

Flow Cytometry: Flavivirus group antigen Antibody (D1-4G2-4-15 (4G2)) - Chimeric [NBP2-52666] - Left: Uninfected cells. Right: Cells infected with yellow fever 17D virus, MOI=1. Image from verified customer review. Analysis using NBP2-52666F conjugate.

Flow Cytometry: Flavivirus group antigen Antibody (D1-4G2-4-15 (4G2)) - Chimeric [NBP2-52666] - Left panel: uninfected VeroB4 cells stained with 4G2 antibody at 1:400. Right panel: VeroB4 cells infected with Yellow-Fever 17D-Venus virus, stained with 4G2 antibody at 1:400. The signal from the 4G2 antibody correlates well with the Venus signal from the virus. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: Flavivirus group antigen Antibody (D1-4G2-4-15 (4G2)) - Chimeric [NBP2-52666] - Detection of Zika virus. Immunofluorescence images of Vero cells infected with ZIKV after 30h infection as well as controls. Cells were fixed in 4% PFA for 30 min at RT. Primary antibody (Rabbit IgG) staining was performed in 0.3% Triton X-100 at 1:20 dilution overnight at 4C, detected using a Goat Anti-Rabbit IgG (1:500) for 1h at RT and counterstained with DAPI.

• Reactivity: V-Vi
• Applications: WB, ELISA, Flow, ICC/IF, IHC, B/N, CyTOF-ready
• Clone: D1-4G2-4-15 (4G2)
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1 mg/ml

Flavivirus group antigen Antibody (D1-4G2-4-15 (4G2)) - Chimeric - Azide and BSA Free Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: This recombinant Flavivirus group antigen Antibody (D1-4G2-4-15 (4G2)) - Chimeric was prepared from Dengue Virus type 2 antigens.
• Specificity: This Flavivirus group antigen Antibody (D1-4G2-4-15 (4G2)) - Chimeric recognises flavivirus group specific antigens (Dengue virus, West Nile Virus, Japanese Encephalitis, Yellow Fever Virus, Zika virus etc). It binds to the fusion loop at the extremity of domain II of protein E.
• Isotype: IgG Kappa
• Clonality: Monoclonal
• Host: Rabbit
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • ELISA 1:100 - 1:2000
  • Flow Cytometry 1:10 - 1:1000
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
  • Neutralization
  • Western Blot 1:100 - 1:2000
• Application Notes: This antibody is CyTOF ready. This chimeric rabbit antibody was made using the variable domain sequences of the original murine IgG2a format, for improved compatibility with existing reagents, assays and techniques. This antibody binds to flavivirus group antigen, protein E. It can be used as an anti-Dengue virus antibody, anti-West Nile virus antibody, anti-Japanese Encephalitis, anti-Yellow Fever Virus or anti-Zika Virus antibody (Aubry et al. 2016) to identify cells infected with these flaviviridae. It binds to the fusion loop at the extremity of domain II of E protein from all four serotypes and prevents syncytia formation (Summers, 1989). The epitope is highly conserved amongst flaviviridae and has been functionally analyzed in detail by Crill and Chang 2004 (PMID: 15564505). Previous studies have used acetone- (Henchal et al. 1982) or methanol-fixed slides (Moreland & Tay, 2010). Please note that binding of this antibody has been reported to be sensitive to reduction and Western Blots should be performed under non-reducing conditions (Lai et al. 2008). A version of 4G2 directly conjugaed to APC was used by Quicke and co-workers (Quicke, 2016) to detect Zika virus in human placental macrophages by flow-cytometry (Quicke 2016). Ramaiah et al. (2016) used NBP2-52709 at a 1:200 dilution to stain for Zika virus in methanol-fixed Vero cells 48 h post-infection.

Reactivity Notes

Dengue Virus, Zika Virus, West Nile Virus, Yellow Fever Virus, Flaviviridae

Packaging, Storage & Formulations

• Storage: Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.
• Buffer: PBS
• Preservative: 0.02% Proclin 300
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for Flavivirus group antigen Antibody (D1-4G2-4-15 (4G2)) - Chimeric - Azide and BSA Free

• Envelope Protein
• Protein E

Background

Flaviviruses are a group of seventy enveloped, positive-sense single stranded RNA viruses belonging to the Flaviviridae family (1-5). Flaviviruses are typically transmitted through arthropods, like mosquitoes and ticks, and can infect humans causing outbreaks and serious illness or death worldwide (1-5). These well-known viral pathogens include dengue virus (DENV), West Nile virus (WNV), Japanese encephalitis virus (JEV), tick-borne encephalitis virus (TBEV), Yellow Fever virus (YFV), and Zika virus (ZIKV) (1-5). Flaviviruses consist of ~10.7 kb RNA genome with one open reading frame (ORF) that is flanked by 5' and 3' untranslated regions (UTRs) (1, 2, 6). The ORF encodes for a polyprotein that is made up of three structural proteins, the capsid (C), premembrane (prM), and envelope (E) proteins, and seven non-structural (NS) proteins (NS1, NS2A/B, NS3, NS4A, 2K, NS4B, and NS5) (1, 2, 5, 6).

The Flavivirus group antigen antibody binds to the fusion loop at the distal end of domain II (EDII) of protein E. Flavivirus E is a glycoprotein that is the main antigen for membrane fusion between virus and host and entry into the cell, as well as inducing protective immunity by stimulating neutralizing antibodies (2, 4, 6). The E protein is 495 amino acids (aa) in length and has a theoretical molecular weight between 55 - 72 kDa, which varies based on glycosylation sites (2). Flavivirus E protein contains three domains (EDI, EDII, and EDIII) followed by a stem that contents to the helix-transmembrane domains (TMDs, TM1 and TM2) (2, 4, 6). The EDII contains an S-S bridge at its distal end and functions as a fusion loop (FL) for virus-mediated membrane fusion and antibody binding (2). Given the important role of Flavivirus protein E in host cell infection and membrane fusion, it has become a target for flavivirus disease prevention and treatment (2, 5).

Alternative names for Flavivirus group antigen include Envelope Protein and Protein E.

References

1. Ngono, A. E., & Shresta, S. (2018). Immune Response to Dengue and Zika. Annual review of immunology. https://doi.org/10.1146/annurev-immunol-042617-053142

2. Rey, F. A., Stiasny, K., Vaney, M. C., Dellarole, M., & Heinz, F. X. (2018). The bright and the dark side of human antibody responses to flaviviruses: lessons for vaccine design. EMBO reports. https://doi.org/10.15252/embr.201745302

3. Zhang, X., Jia, R., Shen, H., Wang, M., Yin, Z., & Cheng, A. (2017). Structures and Functions of the Envelope Glycoprotein in Flavivirus Infections. Viruses. https://doi.org/10.3390/v9110338

4. Laureti, M., Narayanan, D., Rodriguez-Andres, J., Fazakerley, J. K., & Kedzierski, L. (2018). Flavivirus Receptors: Diversity, Identity, and Cell Entry. Frontiers in immunology. https://doi.org/10.3389/fimmu.2018.02180

5. Boldescu, V., Behnam, M., Vasilakis, N., & Klein, C. D. (2017). Broad-spectrum agents for flaviviral infections: dengue, Zika and beyond. Nature reviews. Drug discovery. https://doi.org/10.1038/nrd.2017.33

6. Crill, W. D., & Chang, G. J. (2004). Localization and characterization of flavivirus envelope glycoprotein cross-reactive epitopes. Journal of virology, 78(24), 13975-13986. https://doi.org/10.1128/JVI.78.24.13975-13986.2004

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 Flavivirus group antigen Antibody (NBP2-52666)(11)

Publications using NBP2-52666

• Li Y, Shi S, Xia F et al. Zika virus induces neuronal and vascular degeneration in developing mouse retina Acta neuropathologica communications 2021-05-25 [PMID: 34034828] (IF/IHC)
• Song DH, Garcia G Jr, Situ K et al. Development of a blocker of the universal phosphatidylserine- and phosphatidylethanolamine-dependent viral entry pathways Virology 2021-05-21 [PMID: 34020328] (FLOW)
  
  Details:
  Citation using the Alexa Fluor 647 format of this antibody.
• Rossi F, Josey B, Sayitoglu EC et al. Characterization of zika virus infection of human fetal cardiac mesenchymal stromal cells PloS one 2020-09-17 [PMID: 32941515] (FLOW, Human)
• Thulasi Raman SN, Latreille E, Gao J et al. Dysregulation of Ephrin receptor and PPAR signaling pathways in neural progenitor cells infected by Zika virus Emerg Microbes Infect 2020-12-01 [PMID: 32873194] (IHC-P, Virus)
  
  Details:
  ZIKV
• Cherkashchenko L, Gros N, Trausch A et al. Validation of Flavivirus Infectious Clones Carrying Fluorescent Markers for Antiviral Drug Screening and Replication Studies bioRxiv 2023-04-05 (ICC/IF)
  
  Details:
  Dilutions: 1:1000
• Hans C. Leier, Jules B. Weinstein, Jennifer E. Kyle, Joon-Yong Lee, Lisa M. Bramer, Kelly G. Stratton, Douglas Kempthorne, Aaron R. Navratil, Endale G. Tafesse, Thorsten Hornemann, William B. Messer, Edward A. Dennis, Thomas O. Metz, Eric Barklis, Fikadu G. Tafesse A global lipid map defines a network essential for Zika virus replication Nature Communications 2020-07-21 [PMID: 32694525]
• Martinez Le, Garcia G, Contreras D et Al. Zika Virus Mucosal Infection Provides Protective Immunity J. Virol. 2020-02-12 [PMID: 32051274]
• Garcia G, Paul S, Beshara S et al. Hippo Signaling Pathway Has a Critical Role in Zika Virus Replication and in the Pathogenesis of Neuroinflammation Am. J. Pathol. 2020-02-05 [PMID: 32035058]
• Moreno GK, Newman CM, Koenig MR, Mohns MS Long-term protection of rhesus macaques from Zika virus reinfection J Virol 2019-12-06 [PMID: 31801867]
• Goo L, Debbink K, Kose N et al. A protective human monoclonal antibody targeting the West Nile virus E protein preferentially recognizes mature virions. Nat Microbiol. 2018-11-19 [PMID: 30455471] (WB, Virus)
• Zhang J, Lan Y, Li MY et al. Flaviviruses Exploit the Lipid Droplet Protein AUP1 to Trigger Lipophagy and Drive Virus Production Cell Host Microbe 2018-06-13 [PMID: 29902443] (Virus)