TY - JOUR T1 - Small-scale, high-throughput method for plant N-glycan preparation for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis. A1 - Matsuo,Kouki, Y1 - 2011/02/12/ PY - 2010/11/18/received PY - 2011/02/05/revised PY - 2011/02/08/accepted PY - 2011/2/16/entrez PY - 2011/2/16/pubmed PY - 2011/8/5/medline SP - 200 EP - 2 JF - Analytical biochemistry JO - Anal Biochem VL - 413 IS - 2 N2 - A simple, small-scale, and high-throughput method for preparation of plant N-glycans for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is described. This method entailed the extraction of soluble proteins, pepsin digestion, release of N-glycans by glycopeptidase A, and a three-step chromatographic purification process using cation exchange, anion exchange, and graphitized carbon. Homemade minicolumns using commercially available filter unit devices were used for N-glycan purification steps. All purification steps were designed to be easy. Using this method, N-glycans from 10-mg leaf samples of different plant species and only 2 μg of pure horseradish peroxidase were successfully purified. SN - 1096-0309 UR - https://www.unboundmedicine.com/medline/citation/21320463/Small_scale_high_throughput_method_for_plant_N_glycan_preparation_for_matrix_assisted_laser_desorption/ionization_time_of_flight_mass_spectrometry_analysis_ DB - PRIME DP - Unbound Medicine ER -