Extraction and Verification of Mouse and Human Mucins from Tissue and Fecal Material

Gel-forming mucins are highly O-glycosylated polymeric glycoproteins which have critical roles in tissue protection from environmental insult. To understand their biochemical properties, these samples must be extracted and enriched from biological samples. Here we describe how to extract and semi-purify human and murine mucins from intestinal scrapings or fecal material. As mucins have high molecular weights, traditional gel electrophoresis methods are unable to effectively separate these glycoproteins for analysis. We describe the procedure for making composite sodium dodecyl sulfate urea agarose-polyacrylamide (SDS-UAgPAGE) gels, which allows for accurate verification and band separation of extracted mucins.