Adult Haemonchus contortus were obtained from infected sheep and the N-glycans were prepared using standard laboratory protocols (Paschinger, et al. 2012 (link)) with release of N-glycans using peptide:N-glycosidase A from peptic peptides. After pyridylamination and two rounds of gel filtration, the labelled N-glycans were fractionated by HPLC using an Ascentis® Express 2.7 μ RP-Amide column (150 × 4.6 mm; Sigma-Aldrich)(Yan, et al. 2015 ). Monoisotopic MALDI-TOF MS was performed using a Bruker Autoflex Speed (equipped with a 1000 Hz Smartbeam™-II laser) instrument in positive reflectron mode with 6-aza-2-thiothymine (ATT) as matrix. MS/MS was performed by laser-induced dissociation. Further analysis by MALDI-TOF MS was performed after treatment with either Aspergillus β-galactosidase (Dragosits, et al. 2014 (link)), bovine kidney α-fucosidase (Sigma-Aldrich), α-mannosidases (jack bean from Sigma or Xanthomonas α1,2/3-specific from New England Biolabs), α-galactosidase (coffee bean from Sigma-Aldrich) or recombinant Apis mellifera FDL β-hexosaminidase (Dragosits, et al. 2015 ) in 25 mM ammonium acetate, pH 4.5, at 37 °C overnight (three hours in the case of FDL). For removal of α1,3-fucose or phosphorylcholine residues, selected fractions were dried and incubated for 24 or 48 hours at 0 °C with 3 μl 48% (v/v) hydrofluoric acid prior to evaporation.