Western Blot: Detection of pertussis toxin in culture medium of Bordetella pertussis strain Tohama by Western blotting using anti-pertussis toxin antibody. The toxin consists of five subunits as indicated by S1 to S5.
ELISA: Titration of antibody reactivity of anti-Pertussis antiserum by direct ELISA. Plate was coated with 100μg of pertussis toxin per well and 100μl of the antiserum at the indicated dilution was added to each well and incubated. After washing, goat anti-rabbit-IgG conjugated with HRP was added as secondary antibody. Colour was developed with TMB as substrate.
ELISA: Titration of pertussis toxin by direct ELISA using anti-pertussis toxin antiserum. ELISA plate is coated with indicated amounts of pertussis toxin per well. Antiserum was used at 1/12,500 dilution. ELISA was performed as above. Dynamic range was 100 pg to 10 ng under these conditions.
ANTI-PERTUSSIS TOXIN ANTIBODY
Anti-Pertussis toxin antibody, is a rabbit polyclonal antibody against Bordetella pertussis toxin.
PRODUCT DETAILS – ANTI-PERTUSSIS TOXIN ANTIBODY
- Whole rabbit antiserum with 0.09% sodium azide.
- Immunization was Initiated with toxoid and boosted with native toxin.
- Suitable for Western blotting, ELISA, Dot blotting, Immunoprecipitation, Neutralizing assay.
BACKGROUND
Bordetella pertussis is a Gram-negative, aerobic, pathogenic, encapsulated coccobacillus of the genus Bordetella, and the causative agent of pertussis or whooping cough. B. pertussis is motile and expresses a flagellum-like structure. Its virulence factors include pertussis toxin, adenylate cyclase toxin, filamentous hæmagglutinin, pertactin, fimbria, and tracheal cytotoxin. he bacterium is spread by airborne droplets; its incubation period is 7–10 days on average (range 6–20 days). Humans are the only known reservoir for B. pertussis.
Anti Bordetella pertussis toxin antibody has been developed in response to increasing pressure on the IVD industry to supply highly specific, cost-effective antibody capture systems, which are needed for monitoring vaccination programmes. Perrtussis toxin (PT) is a protein-based AB5-type exotoxin produced by Bordetella pertussis. PT catalyzes the ADP-ribosylation of the α subunits of the heterotrimeric guanine nucleotide regulatory N proteins Gi, Go, and Gt and prevents intracellular signal transduction involving the G proteins. PT consists of one moplecule of each S1 (26 kDa), S2 (22 kDa), S3 (22 kDa), S5 (12 kDa) and two molecule of S4 (12 kDa). The immunogen was highly purified (>90% pure) from Bordetella pertussis strain Tohama by the method of Skelton & Wong. Cytotoxicity of the PT was confirmed by morphological alteration of CHO cells after treatment with 0.1 ng/ml of PT.
REFERENCES
- Alouf JE & Popoff MR (Ed.) The comprehensive Sourcebook of Bacterial Protein Toxins 3rd Ed. Academic Press (2006).
- Skelton SK, Wong KH. Simple, efficient purification of filamentous hemagglutinin and pertussis toxin from Bordetella pertussis by hydrophobic and affinity interaction. J Clin Microbiol. 1990 May;28(5):1062-5.
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