Redox Molecular Signaling Core

项目摘要 科布雷氧化还原生物学和心血管疾病中心的目标是建立一个全国性的 公认的心血管疾病氧化还原生物调节和高级研究和培训计划 促进初级教员科研项目发展，实现重大自主科研经费。 氧化还原分子信号核心(核心C)为Cobre研究人员和受训人员提供访问 用于检测和量化活性氧物种、活性氮物种、 和活性硫物种。此外，该设备还可以分析蛋白质的氧化还原控制。 通过氧化还原蛋白质组和代谢组学发挥作用，为细胞培养缺氧提供集中化的设施 研究并为建立氧化还原信号的细胞培养模型提供专业知识。这一范围的分析 对于路易斯安那州的一家工厂来说，评估氧化还原信号的技术将是无与伦比的。的功能 氧化还原分子信号核心通过两个不同的子核心来完成，即分析氧化还原生物学 亚核和分子信令亚核。分析氧化还原生物子核利用设备进行 高效液相色谱仪、专门的化学发光检测器和质谱仪 光谱(MS)在集中式设施中提供高质量、定量的氧化还原测量， 敬业的技术人员。在第一阶段，该机构购买了Orbitrap Exploris 480质谱仪 这大大扩展了我们对细胞和组织功能进行蛋白质组学分析的能力 氧化还原信号。在第二阶段，我们将通过购买新的 用于活性硫物种定量的设备和额外的质谱学设备以增强 敏感度。分子信令子核心提供了一个集中的设施，用于为分子信号提供帮助 生物学，用于建立氧化还原信号的细胞培养模型，并提供访问关键设备 利用高通量自动毛细管模拟培养中的缺氧细胞损伤和蛋白质分析 电泳法。在第二阶段，我们将通过添加额外的 基于当前Cobre教职员工需求的核心服务，如改进的瞬变方法 原代细胞的转基因和更好地获得下一代测序技术。教职员工和 Redox分子信号核心的工作人员与Cobre研究人员密切合作，以优化实验 设计、执行分析测量、提供核心设备使用方面的培训，并协助处理数据 收集和解读。这些服务为Cobre调查人员提供了宝贵的好处 推进各自的项目，同时加强当地氧化还原生物学研究的基础设施。

PROJECT ABSTRACT The goals of the COBRE Center for Redox Biology and Cardiovascular Disease are to establish a nationally recognized advanced research and training program in redox biology regulation of cardiovascular disease and to facilitate the development of junior faculty research programs to achieve major independent research funding. The Redox Molecular Signaling Core (Core C) provides COBRE investigators and trainees with access to modern equipment for the detection and quantification of reactive oxygen species, reactive nitrogen species, and reactive sulfur species. Additionally, this facility enables the analysis of redox-dependent control of protein function through redox proteomics and metabolomics, provides a centralized facility for cell culture hypoxia studies, and provides expertise for establishing cell culture models of redox signaling. This range of analytical techniques to assess redox signaling will be unmatched for a single facility in Louisiana. The functions of the Redox Molecular Signaling Core are accomplished through two distinct sub-cores, the Analytical Redox Biology Sub-Core and the Molecular Signaling Sub-Core. The Analytical Redox Biology Sub-Core utilizes equipment for high performance liquid chromatography (HPLC), specialized chemiluminescent detectors, and mass spectrometry (MS) to provide high quality, quantitative redox measurements at a centralized facility with dedicated technical staff. During Phase I, the institution purchased an Orbitrap Exploris 480 mass spectrometer that significantly expanded our capability to perform proteomic analysis of cell and tissue function in models of redox signaling. During Phase II, we will expand the functionality of this core through the purchase of new equipment for reactive sulfur species quantification and additional mass spectrometry equipment to enhance sensitivity. The Molecular Signaling Sub-Core offers a centralized facility for providing assistance with molecular biology, for establishing cell culture models of redox signaling, and for affording access to vital equipment for modeling hypoxic cell injury in culture and for protein analysis using high throughput, automated capillary electrophoresis. In Phase II, we will mature and expand the Molecular Signaling Sub-Core by adding additional core services based on the needs of the current COBRE faculty, such as improved methods for transient transfection of primary cells and enhanced access to next generation sequencing technologies. The faculty and staff of the Redox Molecular Signaling Core work closely with the COBRE investigators to optimize experimental design, perform analytical measurements, provide training on the use of core equipment, and assist with data collection and interpretation. These services provide an invaluable benefit to the COBRE investigators to advance their individual projects while enhancing the infrastructure of local redox-biology research.