GLYCOSYL COMPOSTION ANALYSIS OT TWO POMGNT1 SAMPLES (FROM MICE)

两个 POMGNT1 样品（来自小鼠）的糖基组成分析

• 批准号: 7602820
• 负责人: Parastoo Azadi
• 金额: $ 0.12万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602820
• 关键词: Acetylation; Amino Sugars; Blood capillaries; Client; Computer Retrieval of Information on Scientific Projects Database; Detection; Funding; Glycosides; Grant; Hour; Institution; Mass Fragmentography; Methanol; Methods; Monosaccharides; Mus; Procedures; Research; Research Personnel; Resources; Sampling; Silicon Dioxide; Source; United States National Institutes of Health; acetic anhydride; capillary; pyridine

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. Glycosyl composition analysis was performed by combined gas chromatography/mass spectrometry (GC/MS) of the per-O-trimethylsilyl (TMS) derivatives of the monosaccharide methyl glycosides produced from the sample by acidic methanolysis. Methyl glycosides were first prepared from 200 ¿g of the dry sample provided by the client by methanolysis in 1 M HCl in methanol at 80¿C (18-22 hours), followed by re-N-acetylation with pyridine and acetic anhydride in methanol (for detection of amino sugars). The samples were then per-O-trimethylsilylated by treatment with Tri-Sil (Pierce) at 80oC (0.5 hours). These procedures were carried out as previously described (Merkle, R. K. and Poppe, I. (1994) Methods Enzymol. 230:1-15; York W. S., Darvill, A. G., McNeil, M., Stevenson, T. T., and Albersheim, P. (1985) Methods Enzymol. 118:3-40). GC/MS analysis of the TMS methyl glycosides was performed on an HP 5890 GC interfaced to a 5970 MSD, using a Supelco DB-1 fused silica capillary column (30m x 0.25 mm ID).

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。 采用气相色谱/质谱仪(GC/MS)联用技术，对酸解法制备的甲基糖苷的全O-三甲基硅基(TMS)衍生物进行了糖基组成分析。 首先从客户提供的200？g干样品中，在80℃(18-22小时)的1M盐酸中进行甲醇分解，然后在甲醇中与吡啶和乙酸酐进行再次N-乙酰化反应(用于检测氨基糖)。然后在80oC(0.5h)下用Tri-Sill(皮尔斯)处理使样品进行全O-三甲基硅烷化。这些程序如前所述(Merkle，R.K.和Poppe，I.(1994)方法Enzymo.230：1-15；York W.S.，Darvill，A.G.，McNeil，M.，Stevenson，T.和Albersheim，P.(1985)方法。118：3-40)。TMS甲基糖苷的GC/MS分析在HP 5890 GC与5970 MSD接口上进行，使用Supelco DB-1熔融石英毛细管柱(30m×0.25 mm ID)。