title: A procedure for Alcian blue staining of mucins on polyvinylidene difluoride membranes.
abstract: The isolation and characterization of mucins are critically important for obtaining insight into the molecular pathology of various diseases, including cancers and cystic fibrosis. Recently, we developed a novel membrane electrophoretic method, supported molecular matrix electrophoresis (SMME), which separates mucins on a polyvinylidene difluoride (PVDF) membrane impregnated with a hydrophilic polymer. Alcian blue staining is widely used to visualize mucopolysaccharides and acidic mucins on both blotted membranes and SMME membranes; however, this method cannot be used to stain mucins with a low acidic glycan content. Meanwhile, periodic acid-Schiff staining can selectively visualize glycoproteins, including mucins, but is incompatible with glycan analysis, which is indispensable for mucin characterizations. Here we describe a novel staining method, designated succinylation-Alcian blue staining, for visualizing mucins on a PVDF membrane. This method can visualize mucins regardless of the acidic residue content and shows a sensitivity 2-fold higher than that of Pro-Q Emerald 488, a fluorescent periodate Schiff-base stain. Furthermore, we demonstrate the compatibility of this novel staining procedure with glycan analysis using porcine gastric mucin as a model mucin.
authors.affiliation: National Institute of Advanced Industrial Science and Technology (AIST), Open Space Laboratory C-2, 1-1-1 Umezono, Tsukuba, Ibaraki 305-8568, Japan.
authors.forename: Weijie
authors.initials: W
authors.lastname: Dong
authors.forename: Yu-ki
authors.initials: YK
authors.lastname: Matsuno
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kameyama
mesh_terms: D000423:Alcian Blue / Q000032:analysis*; D000818:Animals; D008567:Membranes, Artificial; D009077:Mucins / Q000032:analysis*; D010503:Periodic Acid-Schiff Reaction; D011134:Polysaccharides / Q000032:analysis; D011145:Polyvinyls / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*; D019802:Succinic Acid / Q000737:chemistry*; D013552:Swine; D005465:Fluorocarbon Polymers
chemical_list: D000423:Alcian Blue; D008567:Membranes, Artificial; D009077:Mucins; D011134:Polysaccharides; D011145:Polyvinyls; D019802:Succinic Acid; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers