title: Identification and quantification of mucin expression.
abstract: The major phenotype of CF is the accumulation of mucus, a phenomenon whose relation to the dysfunctional CFTR is still not fully understood. This means that studies of mucus and its main component, the mucins, are important. Due to the large size and high glycosylation level, such questions need special considerations and methodology. We describe methods for the general quantification of heavily glycosylated proteins as the mucins using dot/slot blot. We also describe the separation of the mucins by gel electrophoresis and the identification with specific antibodies on Western blot and by proteomics.
authors.affiliation: Department of Medical Biochemistry, University of Gothenburg, 413 90 Gothenburg, Sweden. kristina.thomsson@medkem.gu.se
authors.forename: Kristina A
authors.initials: KA
authors.lastname: Thomsson
authors.forename: Gunnar C
authors.initials: GC
authors.lastname: Hansson
mesh_terms: D000423:Alcian Blue / Q000032:analysis; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000503:physiopathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D009077:Mucins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D009079:Mucociliary Clearance; D009093:Mucus / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics; D012995:Solubility; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism
chemical_list: D000906:Antibodies; D009077:Mucins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D014357:Trypsin; D000423:Alcian Blue