... byAnthony P. CorfieldThe MucinsGlycoproteinMethodsand Protocols
Isolation of Large Gel-Forming Mucins 33From :Methods in Molecular Biology, Vol. 125: Glycoprotein Methods and Protocols: The MucinsEdited ... Methods in Molecular BiologyTMHUMANA PRESSGlycoproteinMethodsand ProtocolsEdited byAnthony P. CorfieldVOLUME 125The MucinsMethods in Molecular BiologyTMHUMANA ... substi-tution,...
... T-antigen (β-d-Gal 1-3 GalNAc),but also to structures found within the backbone of oligosaccharides (β-d-Gal 1-3 /4GlcNAc) (19). Demonstration of PNA binding is not necessarily evidence of T-anti-gen ... are, however, several other alternatives, such as 3-amino-9-ethylcarbazole (AEC), for use with horseradish peroxidase (HRP); 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium (B...
... of phenol in 70 mL of glycerol.8. Poly-L-lysine-coated slides: slides are immersed in 0.01% (w/v) poly-L-lysine for 5 minand dried at room temperature.3. Methods3 .1. Observation and Measurementof ... A., Allen, A., Morris, E., and Ross-Murphy S. B. (1988) Mucus glycoproteingels: role of glycoprotein polymeric structure and carbohydrate side-chains in gel-forma-tion. Carbohydrate Res. 178,...
... medium byWestern-type dot-blot procedure. Spot aliquots of the homogenates or media on nitrocel-lulose paper, using the dot-blot apparatus and air-dry for 5 min. Perform all ensuing pro-cedures in ... equiv-alent apparatus) to quantify the amount of radioactivity in an identified band on SDS-PAGE.6. Poly- or monoclonal antibodies (IgG-type) against nonglycosylated regions of the mucin-polypep...
... with“naked” cysteine-rich domains (4–7).Histological and biochemical investigations suggested that mucous-producing tis-sues and their secretions contained a complex mixture of mucin-type glycoproteins.However, ... 106) of the gel-formingmucins in particular, there are few separation techniques available to use with these77From: Methods in Molecular Biology, Vol. 125: Glycoprotein Method...
... window with elution profiles of PTH-threonine-GalNAc-Gal (black arrow-heads) and PTH-threonine-GalNAc (gray arrowheads) from the Applied Biosystems Procise. Stan-dard amino acids are shown for comparison ... from Edman degrada-tion: the identification of Xaa-Pro-Xaa-Xaa as a motif for Thr-O-glycosylation. Biochem.Biophys. Res. Commun. 178, 1194–1201.6. Laursen, R. A. (1971) Solid-phase Edman...