Anti-Influenza A Virus Nucleoprotein antibody, mouse monoclonal(C43), pantropic to A type viruses

Product#: 65-110
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Anti-Influenza A Virus Nucleoprotein antibody, mouse monoclonal(C43), pantropic to A type viruses


General information 

Cat. No. 65-110
Size :100 ug
Host Species Mouse
Label Unlabeled
Purification Produced in serum-free medium and purified by proprietary chromatography procedure under mild
    conditions. 90~95% pure by SDS-PAGE
Reactivity Reacts with NP of all influenza A viruses tested, including seasonal H2N2, H3N2, and avian H5N1, H5N2
    and H1N1 (seasonal, pandemic and swine). No cross reactivity with influenza B viruses.
Immunogen Human Influenza A Virus (H2N2) Okada strain
Isotype mouse IgG2A
Application  
  1. Western blotting (300~1,000 fold dilution)
  2. Immuno-precipitation (100 fold dilution)
  3. Immunofluorescent staining (200 fold dilution)
  4. ELISA (assay dependent)
Storage Ship at 4℃, and store at -20℃. Do not freeze
Form 1 mg/ml in PBS, 50% glycerol, filter sterilized
Data Link Swiss-Prot Influenza NP

Description

Influenza virus is an RNA virus, which causes influenza, and belongs to the family Orthomyxoviridae. Influenza virus is classified into three different genera, influenzavirus A, B, and C. They all have similar structures and compositions. The virions are 80-100nm in diameter and usually roughly spherical. The outer surface of the virion is made of a viral envelope containing two major glycoproteins, hemagglutinin (HA) and neuraminidase (NA). Influenzavirus A is further classified into subtypes based on the surface glycoproteins, HA and NA. Currently, there are 16 HA and 9 NA subtypes. The central core of the virion contains the viral RNA genome, which is packaged in the form of ribonucleoprotein complexes.

Influenza virus nucleoprotein (NP) is a major component of the ribonucleoprotein complex and is abundantly expressed during the course of infection. It is a structural protein, which encapsidates the negative strand viral RNA and is essential for RNA transcription, replication and packaging. NP binds the PB1 and PB2 subunits of the viral RNA polymerase and the matrix protein M1, in addition to its binding to ssRNA. NP is also known to interact with variety of other macromolecules of both viral and cellular origins, and these interactions have been shown to be essential for the viral lifecycle.


 
Anti-InfluenzaAVirus-1.png
Fig.1 Immunofluorescence assay of MDCK cells derived from canine kidney cells, and A549 cells derived from human lung carcinoma cells, that were infected with H1N1 influenza virus (A/PuertoRico/8/34).
Samples were taken at 3, 9, and 24 hours post-infection. C43 antibody efficiently detected virus-infected MDCK and A549 cells as early as 3 h after infection. The cells were fixed with 4% paraformaldehyde in phosphate-buffered saline and permeabilized with 0.1% 0.1% Triton X-100 in PBS. The bound antibody was visualized by a further reaction with an Alexa Fluor 488-conjugated secondary antibody
Anti-InfluenzaAVirus-2.png Fig.2 Immunofluorescence assay of 293T cells expressing HA or NP of pandemic (H1N1) 2009 influenza A virus (A/Suita/1/2009). 
C43 specifically recognized NP-expressing cells while a commercially available mouse anti-HA monoclonal 
antibody specifically recognized HA.
65-111_fig1.png
Fig.3 Western blotting of MDCK cells infected with H1N1 (A/PuertoRico/8/34), H5N1 (A/duck/HK/342/78), or H5N2 (A/crow/Kyoto/53/04) using C43 antibody.
Samples were collected at 3, 9, 24, and 48 hours post-infection. C43 detected NP after 3 hours post-infection and detected three different types of influenza viruses. 
Anti-InfluenzaAVirus-3.png Fig.4. Identificationof Influenza Nucleoprotein in crude extract of MDCK cells infected with Influenza A virus (H1N1) PuertoRico/8/34 using C43 mnoclonal antibody. 
10-20% gradient gel,Blotting 15v, 30min (semi-dry)blocking over night, 4℃
1st antibody 1/1000 dilution
2nd antibody 1/10,000 dilution; rabbit polyclonal secodary antibody to mouse IgG- H & L (HRP) (ab97046; abcam). Positions of molecular size markers are shown in kDa on the left. NP size is 56 kDa according to Swiss-Prot.
 

References
This antibody was described and used in the following references.

 

  1. Mizuike R. et al. Development of Two Types of Rapid Diagnostic Test Kits To Detect the Hemagglutinin or Nucleoprotein of the Swine-Origin Pandemic Influenza A Virus H1N1. Clin Vaccine Immunol 18: 494–499 (2011) PubMed ID: 21228147 (IF) 
  2. Ueda M. et al. Maturation efficiency of viral glycoproteins in the ER impacts the production of influenza A virus. Virus Research 136: 91–97 (2008) PubMed ID:18550190 (WB) 
  3. Okuno Y et al . A common neutralizing epitope conserved between the hemagglutinins of influenza A virus H1 and H2 strains. J Virol 67: 2552–2558 (1993) PubMed ID:7682624 (IP) 
  4. Sawaengsak C et al.Intranasal chitosan-DNA vaccines that protect across influenza virus subtypes. Int J Pharm. 2014 Oct 1;473(1-2):113-25. PMID: 24998507 (WB, IF)

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