This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Oligosaccharide Profiling by LTQ Linear Ion Trap Mass Spectrometry (LTQ MS) The N-linked glycans from rhLF were released from the peptide backbone and permethylated as in the previous experiment. The derivatized glycans was diluted with 1mM NaOH in 50% methanol and infused directly into an LTQ Orbitrap Discovery mass spectrometer (Thermo Scientific) at a flow rate of 0.5 ?L/min to obtain a nanospray ionization (NSI). Full mass spectrum of each sample was obtained in the positive ion mode. Fragmentation (MS/MS) of parent ions was performed at an isolation width of 2.0 and collision energy of 30%.
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