Proteomics

蛋白质组学核心通过提供设备和 需要训练有素的人员分析细胞蛋白质组成，蛋白质修饰，蛋白质 定量和蛋白质相互作用。蛋白质组谱分析和蛋白质鉴定服务利用现代 基于质谱计的方法。分析的主要平台是Thermo Finnigan LTQ Line 配备电子转移解离(ETD)的离子陷阱。分离蛋白、凝胶塞和全蛋白质组 分析是受支持的。高丰度样品的制备是通过机器人或人工完成的。 蛋白质、消化和固相萃取(SPE)。各种吸着剂，包括专业吸着剂，如 固相萃取法可用于分离磷酸肽的二氧化钛。米氏H4的纳米级高效液相色谱 平台用于大多数分析，可根据需要提供Triversa Nanomate机器人。数据分析是 使用Mascot、SEQUEST、XITandem和PEAKS算法实现，并进行二次数据分析 脚手架。结果以光盘形式分发，或存放在国际分期付款网络上。 核心通过为蛋白质提供清晰且易于访问的机制来提高研究效率 基于等压标签进行蛋白质的鉴定和相对定量。定量技术 支持的包括cICAT、iTRAQ、TMT和SILAC以及多反应监测(MRM)。分析 同位素标记的样品是使用吉祥物定量程序包实现的。实现了MRM分析 使用TSQ Vantage与Pinpoint和Skyline软件进行实验设计和数据分析。这个 蛋白质组学核心的蛋白质鉴定组件为KCI成员提供了 蛋白质鉴定、蛋白质组学分析和生物标记物鉴定。 核心的蛋白质相互作用组件提供检测的仪器和服务 通过荧光偏振(FP)和表面等离子体共振(SPR)结合蛋白质。这些乐器 在核心生产敏感、准确和实时的蛋白质结合事件测量。因此， 核心的蛋白质相互作用部分支持研究人员提出关于蛋白质-蛋白质的问题 相互作用以及这些相互作用对信号通路和细胞功能的影响。

The Proteomics Core enhances the research productivity of KCI members by providing the equipment and trained personnel necessary for analysis of cellular protein composition, protein modification, protein quantitation and protein interaction. Proteome profiling and protein identification services utilize modern mass spectrometer based methods. The primary platform for analysis is the Thermo Finnigan LTQ Linear lon Trap equipped with Electron Transfer Dissociation (ETD). Isolated protein, gel plug and full proteome analysis are supported. Sample preparation is achieved by robotic or manual depletion of high abundance proteins, digestion and solid phase extraction (SPE). Various sorbents including specialized sorbents such as Ti02 for isolation of phosphopeptides are available for SPE. Nanoflow HPLC from a Michrom H4 platform is utilized for most analyses with a Triversa Nanomate robot available as needed. Data analysis is achieved using Mascot, Sequest, XITandem and PEAKS algorithms with secondary data analysis by Scaffold. Results are distributed as hard copy on CD or by deposition on the international Tranche network. The Core enhances research productivity by providing a clear and easily accessible mechanism for protein identification and for relative quantitation of proteins based on isobaric tags. Quantitation technologies supported include cICAT, ITraq, TMT and SILAC and Multiple Reaction Monitoring (MRM). Analysis of isotopically labeled samples is achieved using the Mascot Quantitation package. MRM analysis is achieved using the TSQ Vantage with PinPoint and Skyline software for experimental design and data analysis. The protein identification component of the Proteomics Core provides KCI members access to technology for protein identification, proteomic profiling and biomarker identification. The protein interactions component of the Core provides instrumentation and services for detection of protein binding by Fluorescence Polarization (FP) and Surface Plasmon Resonance (SPR). The instruments in the Core produce sensitive, accurate and real time measurements of protein binding events. Thus, the protein interactions component of the Core supports investigators in asking questions about protein-protein interactions and the effects of those interactions on signaling pathways and cellular function.