MALDI/TOF-MS OF EIGHT SAMPLES AND ESI-MS ANALYSIS OF ONE SAMPLE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 样品用MALDI/TOF-MS和ESI-MS分析。具体步骤如下所示。 基质辅助激光解吸飞行时间质谱仪(MALDI-TOF) MALDI/TOF-MS色谱柱为反射镜正离子模式，以二羟基苯甲酸(DHBA，20 mg/mL甲醇：水)为基质。使用4700蛋白质组学分析仪(应用生物系统)获得所有光谱。 电喷雾电离线性离子陷阱质谱仪(ESI-LCQ/MSN) 采用LC Q-MS(Thermo Finnigan)，用注射器泵(哈佛仪器)将溶解于1 mM氢氧化钠中的过甲基化多聚糖(5pmol/L)以1？L/分钟的恒定流速直接输入LC-Q仪器进行质量分析。毛细管温度设置为200℃，正离子模式下进行MS分析。在MS/MS中，碰撞能被设置为35%的碎裂。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The samples were analyzed by MALDI/TOF- MS and ESI-MS. The procedures are shown in detail below. Matrix-assisted laser-desorption time-of-flight mass spectrometry (MALDI-TOF) MALDI/TOF-MS was performed in the reflector positive ion mode using ¿-dihyroxybenzoic acid (DHBA, 20mg/mL solution in 50%methanol:water) as a matrix. All spectra were obtained by using a 4700 Proteomics analyzer (Applied Biosystems). ElectroSpray Ionization  Linear Ion Trap mass Spectrometry (ESI-LCQ/MSn) Mass analysis by LCQ-MS (Thermo Finnigan) was performed by direct infusion of permethylated glycans dissolved in 1 mM NaOH in 50% methanol ( 5 pmol/¿L) into the LCQ instrument at a constant flow rate of 1 ¿L/min via a syringe pump (Harvard Apparatus). The capillary temperature was set to 200oC and MS analysis was performed in positive ion mode. The collision energy was set at 35% for fragmentation in MS/MS.