CORE--PROTEOMICS

核心--蛋白质组学

• 批准号: 6972210
• 负责人: THOMAS C VANAMAN
• 金额: $ 13.8万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6972210
• 关键词: SDS polyacrylamide gel electrophoresis; biomedical facility; computer program /software; high performance liquid chromatography; immunocytochemistry; mass spectrometry; matrix assisted laser desorption ionization; posttranslational modifications; protein quantitation /detection; protein sequence; proteomics; robotics; two dimensional gel electrophoresis

The proteomics core is designed to provide quantitative and qualitative protein profiling and identification analysis, as well as post-translational modification and sequence analysis through a combination of 2D gel, mass spectrometry and Edman sequencing analysis services. The core is equipped with Bio-Rad IPG and SDS-PAGE apparatus for 2D gel analysis, a Typhoon phosphorimager/scanner system for detection and quantitation of fluorescent protein dyes (Cy-dyes) and stains (SYPRO-Ruby, Pro-Q Diamond) used to analyze 1D and 2D gels, a Ciphergen SELDI-TOF protein profiling system, an ABI QSTAR II Q-TOF Mass Spectrometer with both MALDI and LC/ESI source capabilities, an ABI Procise 4 column Edman pulsed liquid phase Sequencer, Waters HPLC systems and multiple PCs for data acquisition and analysis including a site license for PDQuest software and a 5 station site license for Ciphergen software. The fully trained technical staff identified below will assist investigators with the full range of proteomics analytical services. The staff will instruct investigators and their laboratory staff (students, post-doctoral fellows and technicians) in sample preparation, 2D gel analysis and quantitation by fluorescent staining methods. Protein fragmentation, mass peptide mapping and protein sequencing by both MS/MS mass spectrometry and chemical sequencing will be performed by Dr. Beach on samples provided by individual investigators. Preparation and analysis of samples with the Ciphergen system will be facilitated by automated robotic work stations operated under the supervision of Dr. Beach. Data analysis will be performed by investigators and their colleagues through multiple distant work stations connected to the Ciphergen system via the web.

蛋白质组学核心旨在提供定量和定性蛋白质分析和鉴定分析，以及通过2D凝胶，质谱和Edman测序分析服务的组合进行翻译后修饰和序列分析。核心配备了用于2D凝胶分析的Bio-Rad IPG和SDS-PAGE装置，用于检测和定量荧光蛋白染料（Cy-dyes）和染色剂的Typhoon磷光成像仪/扫描仪系统（SYPRO-Ruby，Pro-Q Diamond），Ciphergen SELDI-TOF蛋白质谱分析系统，具有MALDI和LC/ESI源功能的ABI QSTAR II Q-TOF质谱仪，ABI Procise 4柱Edman脉冲液相测序仪、沃茨HPLC系统和用于数据采集和分析的多台PC，包括 PDQuest软件的站点许可证和Ciphergen软件的5站站点许可证。下面确定的经过全面培训的技术人员将协助研究人员提供全方位的蛋白质组学分析服务。工作人员将指导研究人员及其实验室工作人员（学生，博士后研究员和技术人员）进行样品制备，2D凝胶分析和荧光染色方法定量。由Dr.海滩对个体研究者提供的样本进行蛋白质片段化、质量肽图谱分析以及通过MS/MS质谱法和化学测序法进行蛋白质测序。自动化机器人工作将有助于用Ciphergen系统制备和分析样品 在海滩博士的监督下运作的车站。数据分析将由研究者及其同事通过网络连接到Ciphergen系统的多个远程工作站进行。