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. 2023:2657:197-205.
doi: 10.1007/978-1-0716-3151-5_14.

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

Mackenzie Melvin  1 , Noah Fancy  1 , Darrek Kniffen  1 , Kirk Bergstrom  2
Affiliations

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

Mackenzie Melvin et al. Methods Mol Biol. 2023.

Abstract

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.

Keywords: Alkalization; Composite gels; Dialysis; Fecal; Gel electrophoresis; Glycan; Glycoprotein; Goblet cell; Intestine; Mucin; Mucin extraction; Reduction.

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References

    1. Michael AM, David JT, Jeffrey AW (2015) Chapter 14 – Mucins and mucus. In: Jiri M, Warren S, Michael WR, Brian LK, Hilde C, Bart NL (eds) Mucosal immunology, 4th edn. Academic, Boston, pp 231–250. https://doi.org/10.1016/B978-0-12-415847-4.00014-8 - DOI
    1. Malin EVJ, Gunnar CH (2016) The mucins. In: Michael JHR (ed) Encyclopedia of immunobiology. Academic, Oxford, pp 381–388. https://doi.org/10.1016/B978-0-12-374279-7.02019-1 - DOI
    1. Godl K, Johansson ME, Lidell ME, Morgelin M, Karlsson H, Olson FJ et al (2002) The N terminus of the MUC2 mucin forms trimers that are held together within a trypsin-resistant core fragment. J Biol Chem 277(49):47248–47256. https://doi.org/10.1074/jbc.M208483200 - DOI - PubMed
    1. Carlstedt I, Herrmann A, Hovenberg H, Lindell G, Nordman H, Wickstrom C et al (1995) ‘Soluble’ and ‘insoluble’ mucins – identification of distinct populations. Biochem Soc Trans 23(4):845–851. https://doi.org/10.1042/bst0230845 - DOI - PubMed
    1. Johansson ME, Hansson GC (2012) Analysis of assembly of secreted mucins. Methods Mol Biol 842:109–121. https://doi.org/10.1007/978-1-61779-513-8_6 - DOI - PubMed

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