Methods to Assess the Direct Interaction of C. jejuni with Mucins

Methods Mol Biol. 2017:1512:107-115. doi: 10.1007/978-1-4939-6536-6_10.

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.

Keywords: Campylobacter jejuni; Mucin array; Mucus; PVDF; SDS PAGE; Slot blot.

MeSH terms

  • Animals
  • Avian Proteins / chemistry
  • Avian Proteins / metabolism*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism*
  • Biotin / chemistry
  • Blotting, Far-Western / methods*
  • Campylobacter jejuni / chemistry
  • Campylobacter jejuni / metabolism*
  • Chickens
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Dyes / chemistry
  • High-Throughput Screening Assays*
  • Humans
  • Mucins / chemistry
  • Mucins / metabolism*
  • Protein Array Analysis
  • Protein Binding
  • Protein Interaction Mapping
  • Staining and Labeling / methods

Substances

  • Avian Proteins
  • Bacterial Proteins
  • Fluorescent Dyes
  • Mucins
  • Biotin