fetuin after denaturation  (New England Biolabs)

Journal: Scientific Reports

Article Title: High-throughput analysis of N-glycans using AutoTip via glycoprotein immobilization

doi: 10.1038/s41598-017-10487-8

Figure Lengend Snippet: Rate of glycoprotein immobilization and PNGase F release of N-glycans. ( a ) Fetuin was conjugated to Aminolink resin via reductive amination, whereas the unbound proteins in supernatant were measured by a BCA assay. Fetuin was quickly immobilized to the resin within 4 h; ( b ) N-glycans were released by PNGase F in 25 mM ammonium bicarbonate at 37 °C. The PNGase F digestion was complete within 2 h. Four measurement were made for each condition and error bar represented standard deviation.

Article Snippet: Each AutoTip conjugated 20 µg of fetuin after denaturation (88 µL fetuin protein in HPLC water +10 µL denaturing buffer (NEB); 100 °C/10 min).

Techniques: BIA-KA, Standard Deviation

Journal: Scientific Reports

Article Title: High-throughput analysis of N-glycans using AutoTip via glycoprotein immobilization

doi: 10.1038/s41598-017-10487-8

Figure Lengend Snippet: Modification of sialic acids using carbodiimide coupling on AutoTip. Fetuin protein was modified by p-Toluidine in the presence of EDC after immobilization on the beads (Four AutoTips). Sialylated N-glycan, S2H5N4, was used to monitor completion of p-Toluidine-EDC reaction. Without derivatization, MALDI-MS detected no sialic acid H5N4 ( a ) and two sialic acids S2H5N4 ( c ); For incomplete derivatization, MALDI-MS detected ( a ), ( c ), and p-Toluidine labeled two sialic acids S2H5N4 ( b ); When completely labeled, only ( b ) was detected by MALDI-MS.

Article Snippet: Each AutoTip conjugated 20 µg of fetuin after denaturation (88 µL fetuin protein in HPLC water +10 µL denaturing buffer (NEB); 100 °C/10 min).

Techniques: Modification, Labeling