Anti-Influenza A Virus NP antibody, mouse monoclonal (C43)
Product code65-110Size

100 µgStorage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from hybridoma cell culture medium

Immunogen

Human Influenza A Virus (H2N2) Okada strain

Isotypemouse IgG2aκ

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.

Application1.Western blotting (300~1,000 fold dilution)
2. Immunoprecipitation (100 fold dilution)
3. Immunofluorescent staining (200 fold dilution)
4. ELISA (assay dependent)

Background

Influenza virus is an RNA virus, which causes influenza, and belongs to thefamily Orthomyxoviridae. Influenza virus is classified into three different
genera, influenzavirus A, B, and C. They all have similar structures andcompositions. 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) isa 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 subunitsof 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 othermacromolecules of both viral and cellular origins, and these interactions havebeen shown to be essential for the viral lifecycle.

Data Link

UniProtKB: Influenza NP

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 65-110 Anti-Influenza A Virus NP antibody, mouse monoclonal (C43)

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 virusinfected 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% Triton X-100 in PBS.
The bound antibody was visualized by a further reaction with an Alexa Fluor 488-conjugatedsecondary antibody.

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.
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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 as a primary antibody.
Samples were collected at 3, 9, 24, and 48 hours post-infection. C43 detected NP after 3 hours postinfection and detected three different types of influenza viruses.

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, 60min (semi-dry)

Blocking

overnight, 4℃

1st antibody

1/1000 dilution

2nd antibody

1/10000 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 product has been used in the following publication
1.Mizuike R. et al. Development of Two Types of Rapid Diagnostic Test Kits To Detect theHemagglutinin 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
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BioAcademia,Inc.

Tel. 81-6-6877-2335

Fax. 81-6-6877-2336

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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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