Anti-Helicobacter pylori flagellin antibody, mouse monoclonal (hp-01)
Product code64-108Size

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

Crude extract of Helicobacter pylori

Isotype

Mouse IgG1κ

Reactivity

Helicobacter pylori flagellin (FlaA and FlaB) protein

Special notes

N/A

Application

1. Western blotting (1/500~1/1,000)
2. ELISA (assay dependent)
Other applications have not been tested.

Background

Helicobacter pylori (H. pylori) is a gram-negative, spiral-shaped microaerophilic
bacterium which is linked to the development of duodenal ulcers and stomach
cancer. H. pylori strains produce important virulence factor. Motility isconferred to the bacteria by several sheathed flagella and is regarded as one of
their principal virulence factors. Characterization of the proteins associatedwith the flagella components indicate the hook protein (FlgE) is 78 kDa in size
while the filament consists of two flagellin proteins, FlaA and FlaB. The H.

pylori flaA and flaB genes both coded for proteins with 510 and 514 amino acids
and molecular masses of 53 kDa and 54 kDa, respectively.

Both flagellins

share considerable amino acid homology (58% identity).

Data Link

UniProt KB:
P0A0S1 (FLAA/HELPT), Flagellin A,
Q07911 (FLAB/HELPT) ,Flagellin B

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Fig.1. Detection of polypeptide of H. pylori by Western blotting with MAb (hp-01).
1: Crude extract of H. pylori (isolated strains)
2: Crude extract of H.pylori (JCM12093)
3: Crude extract of Campylobacter jejuni
4: Crude extract of Campylobacter coli
5: Crude extract of Escherichia coli (E.coli)
The hp-01 antibody was used at 1/500 dilution. The HRPO-conjugate goat anti-mouse IgG wasused at 1/4,000 as the second antibody and visualized by DAB (3,3’-Diaminobenzidine). The hp-01
reacted with 53 kDa protein in the extract of H. pylori, but did not react with the extracts ofCampylobacter spp. and E. coli. A protein band 53 kDa in size corresponds to the expected size ofthe flagellin.

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Fig.2. Detection of FlaA and FlaB protein ofH. pylori by Western blotting with hp-01
1: Crude extract of H.pylori (JCM12093)
2: Recombinant FlaA protein expressed in E. coli
3: Recombinant FlaB protein expressed in E. coli
4: Negative control (BL21(DE3))
The hp-01 antibody was used at 1/500 dilution. The HRPO-conjugate goat anti-mouse IgG was used
at 1/4,000 as the second antibody and visualized by Chemi-Luminescence. The hp-01 reacted withrecombinant FlaA and FlaB proteins expressed in E. coli.

Fig.3. Titration of flagellin in the extract of H. pylori cells by indirect ELISA using hp-01.
The wells of plate were coated indicated amountsof the extract of H. pylori cells per well. After blocking with 5% skim milk, 100 µl of hp-01 at 1/500
dilution was added to the each well. The HRPO-conjugated goat anti-mouse IgG (100μl, x2000
dilution) was added. Color was developed with OPD (orthophenylenediamine) as substrate. Opticaldensity (OD) measured at 490nm.

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The hp-01 reacted with a standard strain of H. pylori (JCM12093) and isolated strains, such as
England strain, USA strains (MR31 and RD26 strains). However hp-01 did not react with other foodpoisoning bacteria such as Campylobacter jejuni, Campylobacter coli and enterotoxigenic Escherichia

coli (ETEC).
Reference: There has been no publication using this antibody.
Please let us know when your research using this antibody is published. We will offer one vial of ourantibody as compliment.

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