title: Methods to Assess the Direct Interaction of C. jejuni with Mucins.
abstract: Studies of the interaction of bacteria with mucus-secreting cells can be complemented at a more mechanistic level by exploring the interaction of bacteria with purified mucins. Here we describe a far Western blotting approach to show how C. jejuni proteins separated by SDS PAGE and transferred to a membrane or slot blotted directly onto a membrane can be probed using biotinylated mucin. In addition we describe the use of novel mucin microarrays to assess bacterial interactions with mucins in a high-throughput manner.
authors.affiliation: Conway Institute, School of Medicineand Medical Science, University College Dublin, Dublin, Ireland.
authors.forename: Marguerite
authors.initials: M
authors.lastname: Clyne
authors.affiliation: Conway Institute, School of Medicineand Medical Science, University College Dublin, Dublin, Ireland.
authors.forename: Gina
authors.initials: G
authors.lastname: Duggan
authors.affiliation: Conway Institute, School of Medicine, University College Dublin, Dublin, Ireland.
authors.forename: Julie
authors.initials: J
authors.lastname: Naughton
authors.affiliation: Conway Institute, School of Medicineand Medical Science, University College Dublin, Dublin, Ireland. billy.bourke@ucd.ie.
authors.forename: Billy
authors.initials: B
authors.lastname: Bourke
mesh_terms: D000818:Animals; D030161:Avian Proteins / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001710:Biotin / Q000737:chemistry; D026481:Blotting, Far-Western / Q000379:methods*; D016123:Campylobacter jejuni / Q000737:chemistry / Q000378:metabolism*; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000737:chemistry; D057166:High-Throughput Screening Assays; D006801:Humans; D009077:Mucins / Q000737:chemistry / Q000378:metabolism*; D040081:Protein Array Analysis; D011485:Protein Binding; D025941:Protein Interaction Mapping; D013194:Staining and Labeling / Q000379:methods
chemical_list: D030161:Avian Proteins; D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D009077:Mucins; D001710:Biotin
keywords: Campylobacter jejuni; Mucin array; Mucus; PVDF; SDS PAGE; Slot blot