Thermo Orbitrap Eclipse Tribrid with ETD and an Ultimate 3000 RSLCnano System

该提案正在为带有ETD的Thermo Orbitrap Eclipse Tribrid和Ultimate 3000 RSLCnano寻求资金 系统，以满足来自卡内基两个部门的10个实验室对蛋白质组学分析日益增长的需求 机构（九个由NIH资助）和斯坦福大学的三个实验室（两个由NIH资助）。CISMS 配备一台仪器Orbitrap Q Exactive HF。由于只有HCD模式而缺少MS 3功能， 由于缺乏成熟的组态和软件，现有的仪器已不能满足 目前NIH资助的研究。特别是，许多研究人员在翻译后领域的需求， 修饰（如O-GlcNAc酰化、O-岩藻糖基化、异戊二烯化、甲基化）、交联 质谱，细胞类型特异性和单细胞蛋白质组学，rRNA修饰，蛋白质-RNA界面 映射和使用TMT多路复用的增强量化，目前没有满足现有的 仪器仪表因此，计划收购Orbitrap Eclipse ETD对于开发和 完成NIH支持的研究，从而得到我们研究界的全力支持。拟议 仪器将安置在卡内基科学研究所质谱设施（CIS-MS）内， 被整个地区的研究人员广泛使用。要求的Orbitrap Eclipse与ETD不仅翻了一番 该设施的能力，但也提供了几个关键的能力，目前的文书， 设施。特别是，ETD和混合EThcd碎片模式，MS 3和复杂的 配置和软件，是必不可少的项目，研究位点特异性翻译后修饰 分析，如O-GlcNAc酰化和O-岩藻糖基化，和异戊二烯化，以及使用MS可裂解的结构分析 交联剂和rRNA转录后修饰。执行实时搜索的能力， 同步前体选择（SPS）和MS 3片段化用于TMT同量异位素定量的要求来自 几个小组的项目，要求高准确性，高灵敏度和高通量的蛋白质组学和 磷酸化蛋白质组学、细胞类型蛋白质组学和单细胞蛋白质组学。运营和维护 仪器将由CIS-MS经验丰富的工作人员进行。卡内基机构提供了567，739.80美元 匹配所需的，以满足系统的全部价格，并确保充分支持设施主任和两个 支持人员，以及第2年的服务合同和仪器的剩余有效寿命 一年保修期。

This proposal is seeking funds for a Thermo Orbitrap Eclipse Tribrid with ETD and an Ultimate 3000 RSLCnano system to meet increasing demands for proteomic analysis for 10 labs from two departments of Carnegie Institution (nine are funded by NIH) and for three labs at Stanford University ( two are funded by NIH). The CISMS is equipped with one instrument Orbitrap Q Exactive HF. With only HCD mode and lack of MS3 functionality, lack of sophisticated configuration and software, the current instrument can no longer meet the demands of current NIH-funded research at our institution. In particular, the needs of many researchers in the area of posttranslational modifications (such as O-GlcNAcylation, O-fucosylation, prenylation, methylation), cross-linking mass spectrometry, cell-type specific and single-cell proteomics, rRNA modification, protein-RNA interface mapping, and enhanced quantification using TMT multiplexing, are currently not being met with existing instrumentation. Thus, the planned acquisition of Orbitrap Eclipse ETD is crucial for the development and completion of NIH-supported research, and thereby is fully backed by our research community. The proposed instrument is to be housed within Carnegie Institution for Science Mass Spectrometry facility(CIS-MS) which is highly used by researchers throughout the region. The requested Orbitrap Eclipse with ETD not only doubles the capacity of the facility, but also provides several key capabilities not available on the current instrument of the facility. In particular, ETD and hybrid EThcd fragmentation modes, with MS3 and with sophisticated configuration and software, are essential for projects that study site-specific post-translational modification analysis such as O-GlcNAcylation and O-fucosylation, and prenylation, and structural analysis using MScleavable cross-linker, and rRNA post-transcriptional modifications. The ability to perform real-time search, synchronous precursor selection (SPS) and MS3 fragmentation for TMT isobaric quantification is requested from several groups for projects that demand high accuracy, high sensitivity and high throughput for proteomics and phosphoproteomics, cell type proteomics and single-cell proteomics. Operation and maintenance of the instrument will be performed by experienced staff in CIS-MS. The Carnegie institution provides the $567,739.80 match required to meet the full price of the system, and ensures full support for the facility director and two supporting staff, as well as service contract for year 2 and the remaining effective lifetime of the instrument after the one-year warranty.