AS01 020 | Clonality: Polyclonal | Host: Hen | Reactivity: Helicobater pylori
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whole cells of Helicobacter pylori, standard laboratory strain (17-1 e.g. CCUG 17875) emulsified with an adjuvant.
Panel 1: secondary antibody only
A. - Hoechst, B. - anti- chicken Alexa 568, dilution 1: 500, C. - Hoechst and anti-chicken Alexa 568
Panel 2: primary and secondary antibody detection
A. - Hoechst, B. - chicken anti-Helicobacter pylori at 1: 200 and anti-chicken Alexa 568, dilution 1: 500, C. - Hoechst, chicken anti-Helicobacter pylori at 1: 200 and anti-chicken Alexa 568
Protocol Immunofluorescence for Helicobacter pylori cells:
Harvest Helicobacter pylori from blood agar plates in 0.9% NaCl Put onto coverslips in 24-well plate and spin down at 1500g for 5min Fix with 4% Paraformaldehyde in PBS Wash with PBS 5 min triton X-100 0.2% in PBS Wash with PBS
Block with 4% foetal calf serum in PBS
Chicken anti-Helicobacter pylori primary antibody (Agrisera cat# AS01 020) diluted in 4% FSC in PBS 1 hour at room temp
Wash with PBS
Goat anti-Chicken Alexa fluor 568 (Invitrogen cat# A11041) in PBS 30 min at room temperature
Wash with PBS and water
3 min Hoechst 1ug/ml in water
Wash in water
Courtesy Dr. Phebe Verbrugghe, QEII Medical Centre, Australia
IgY concentration is 23.5 mg/ml
Western blot has been done using whole extracts of Helicobacter pylori. Antibody recognised several low and high molecular weight H. pylori proteins. Some of them are from the surface of bacteria.
Helicobacter pylori is nowadays recognised as one of the most common human pathogens, and has been recently defined as a class 1 carcinogen. It has an ability to colonise gastric mucosa, where it causes chronic gastritis and is a major cause for duodenal and sometimes gastric ulcers.
Lertsethtakarn et al. (2015). Helicobacter pylori CheZHP and ChePep form a novel chemotaxis-regulatory complex distinct from the core chemotaxis signaling proteins and the flagellar motor. Mol Microbiol. 2015 Jun 10. doi: 10.1111/mmi.13086.