CORE--PROTEIN MICROSEQUENCING

描述：蛋白质微测序共享资源的目的是 在蛋白质测序和蛋白质合成方面提供最先进的方法 分析.蛋白质测序被描述为一种独特的技术， 需要特定的、专用的和专门的仪器。工作人员 包括核心负责人、一名技术人员和两名助手。测序完成 在贝克曼-伯顿测序仪上，使用气相化学。的氨基酸 使用Pico-Tag化学进行分析。用于凝胶内和凝胶外的方案 印迹上的胰蛋白酶消化酶由该机构的工作人员开发。 研究者可以使用设施上描述的协议？S 主页或消化和分离所得肽片段， 反相HPLC系统。HPLC级分中存在的肽为 在位于UCLA的灵敏MALDI质谱分光光度计上分析 质谱分析中心。该仪器于1994年作为联合 蛋白质微测序共享资源和加州大学洛杉矶分校中心的努力， 质谱，大部分资金来自CCSG。 该设施的具体目标包括：1）最先进的 蛋白质微测序的方法学; 2） 敏感的氨基酸分析; 3）协助和信息实验室 蛋白质结构测定的研究者; 4）其他蛋白质 服务，包括蛋白质裂解，质量分析和分离， 通过凝胶方法的HPLC测定内部片段。 该设施的经营政策包括确定优先事项 的样本进行测序。最优先考虑的是 癌症中心和生物系的研究人员 化学，为设施提供空间，然后给其他成员 UCLA社区。

Description: The purpose of the Protein Microsequencing Shared Resource is to provide state-of-the-art methodology in protein sequencing and protein analysis. Protein sequencing is described as being a unique technology, requiring specific, dedicated, and specialized instrumentation. The staff includes the Core Leader, a technician, and two helpers. Sequencing is done on a Beckman-Porton Sequencer, using gas phase chemistry. The amino acid analyses are done using Pico-Tag chemistry. The protocols for in-gel and on-blot tryptic digestions were developed by the staff of the facility. Investigators can either use the protocols described on the facility?s homepage or the digestion and separation of the resulting peptide fragments on a reverse phase HPLC system. The peptides present in the HPLC fractions are analyzed on a sensitive MALDI mass spectrophotometer located in the UCLA Center for Mass Spectrometry. This instrument was acquired in 1994 as a joint effort of the Protein Microsequencing Shared Resource and the UCLA Center for Mass Spectrometry, with a major portion of the funding from the CCSG. The specific goals of the facility include providing: 1) state-of-the-art methodology in protein microsequencing to the UCLA research community; 2) sensitive amino acid analyses; 3) assistance and information to laboratory investigators about determination of protein structure; and 4) other protein services, including protein cleavages, mass analyses, and isolation of internal fragments by HPLC of gel methods. The operating policies of the facility include a determination of the priority of samples received for sequencing. The highest priority goes to members of the Cancer Center and to researchers in the Department of Biological Chemistry, which provides space for the facility, and then to other members of the UCLA community.