This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Methods: The N-glycans extracted from the sample (1317046), which were analyzed earlier by MALDI/TOF- MS and ESI-MS/MS (LCQ-MS) have been reported were analyzed by Nanospary ionization mass spectrometry (NSI-MS) using a LTQ-MS (Thermo Finnigan).NSI-MS is more sensitive in detecting the signals for glycans more than ESI-MS. More reliable information about their structure can be obtained from NSI-MS/MS spectrum than either ESI-MS or MALDI MS spectrum.NSI-MS analysis was done with the following conditions. NanoelectroSpray Ionization Linear Ion Trap mass Spectrometry (ESI-LTQ/MSn)Mass analysis by LTQ-MS (Thermo Finnigan) was performed by direct infusion of permethylated glycans dissolved in 1 mM NaOH in 50% methanol (5 pmol/ L) into the LTQ instrument using a nanoelectrostray source at syringe flow rate of 0.40 L/min. The capillary temperature was set to 210oC and MS analysis was performed in positive ion mode. The collision energy was set at 28% for fragmentation in MS/MS.
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