Anti-Influenza B Virus HA antibody, mouse monoclonal (1H12)
Product code65-160Size

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 B Virus strain Nagasaki/1/87, one of the strains of B/Victoriagroup.

Isotype

Mouse IgG1κ

Reactivity

According to Ref.1 during epidemic in Osaka 1996-97, 1H12 antibody reactedwith HA protein of most of Influenza B virus isolates belonging to Victoria grouptested (70/73 strains) and none of clinical 27 isolates belonging to Yamagata
group as examined by PAP staining. However later test with IF staining it also
reacts with some of Yamagata group strains like B/Florida/4/2006. However,note that HA changes during passages and may change reactivity to thisantibody. No cross reactivity with any strains of influenza A virus.

Application

1. Western blotting (1/500~1/1,000 dilution)
2. Immunofluorescent and Immunocytochemical staining (1/100~1/200dilution)
3. Immunoprecipitation (1/200 dilution)
4. ELISA (assay dependent)

BackgroundHemagglutinin (HA) binds to sialic acid-containing receptors on the cellsurface, bringing about the attachment of the virus particle to the cell.
Plays a major role in the determination of host range restriction andvirulence. Class I viral fusion protein. Responsible for penetration of thevirus into the cell cytoplasm by mediating the fusion of the membrane of
the endocytosed virus particle with the endosomal membrane. Low pH inendosomes induce an irreversible conformational change in HA2, releasing
the fusion hydrophobic peptide. Several trimers are required to form acompetent fusion pore.
Post-translational modification
HAo is consists of 584 amino acids with molecular mass of 63,275. In naturalinfection, inactive HA is matured into HA1 and HA2. By sequence similarity it
is indicated to be palmitoylated.

Data Link

UniProtKB P03460 Influenza B/Lee/1940 HA protein.

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: 65-160 Anti-Influenza B Virus HA antibody, mouse monoclonal (1H12)

Fig. 1 Detection of HA protein in the crude extracts of MDCK cells infected with various Influenza B
virus strains by western blotting using 1H12 monoclonal antibody.
1. B/Mie/1/1993
2. B/Johannes Burg/5/1999
3. B/Florida/4/2006
4. B/Lee/1940
5. B/Florida/4/2006
6. Shandong/7/97
7. B/Malasia/2506/2004
8. B/Massachusetts/2/2012
First antibody was used at 1/500 dilution and as 2nd antibody, HRP-conjugated goat anti-mouse IgG
antibody was used at 1/10,000 dilution. Positions of marker proteins are indicated in kDa on the left. Clone
1H12 recognizes an epitope on HA2 region.

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Fig.2

Immunofluorescence assay of MDCK (canine kidney ) cells infected with Influenza B virus, using

anti-Influenza B virus HA antibody (clone 1H12).
Anti-Influenza B Virus HA antibody (clone 1H12) efficiently detected the viruses in the infected MDCK
cells with B/Malaysia/2506/2004 and B/Florida/4/2006 virus strains. The cells were ﬁxed with 4%paraformaldehyde in phosphate-buffered saline (PBS) and permeabilized with 0.1% Triton X-100 in PBS.
The bound antibody was visualized by a further reaction with an Alexa Fluor 488-conjugated secondary
antibody (green).
Image on the left is a negative control, mock-infected MDCK cells.

References: This antibody was described and used in the following reference.
1.Nakagawa N. et al. Rapid detection and identification of two lineages of influenza
B strains with monoclonal antibodies. J Virol Methods. 1999;79:113-2 ICC, IP

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