Skin Proteomics Core

UAB皮肤病研究中心的蛋白质组学核心D为试点/可行性获奖者提供支持， SDRC相关研究人员和其他皮肤病研究人员。这种支持包括协商 由两名经验丰富的高级教师组成，他们在二维凝胶蛋白质组学和蛋白质质谱学方面具有专长， 由一名精通多种蛋白质组学/质谱分析方法的初级教员提供实践支持 并应用于皮肤病的研究，培训所提供的核心技术，获得 仪器和研讨会由外部专家在蛋白质组学和质谱。的具体服务 核心内容包括：（1）开发实验设计和统计考虑， 实验，包括蛋白质组学和质谱组成部分;（2）蛋白质的二维凝胶分离 从组织、细胞、亚细胞部分和特定体液（血浆/血清、尿、汗、眼泪、组织 间质和水疱液）-这涉及等电聚焦/SDS-PAGE或透明天然凝胶电泳 并利用Cy2、Cy3和Cy5荧光染料和Sypro红宝石红后染色、成像和成像 使用DeCyder软件进行分析，机器人凝胶点拾取，化学处理和蛋白酶消化，以及 肽质量指纹图谱;（3）膜相关蛋白的1D-SDS-PAGE凝胶分离， nanoLC-ESI-串联质谱法;（4）同位素标记方法（iTRAQ试剂）进行 定量分析处理和时间对组织、细胞和液体蛋白质组成员的影响; 和（5）开发特定的多反应离子监测（MRM）质谱法， 定量肽，允许定量分析个别，指定的蛋白质。此外，LC-MRM-MS 方法可用于分析皮肤病研究中使用的试剂，例如绿色茶 多酚和葡萄籽提取物，以及它们在血液和其他液体中的药物代谢物。一个特殊 重点是保持方法的质量控制。实验室信息管理系统 将被用来监督核心的运作将对研究者进行单独的核心使用培训 技术以及与其他NIH资助的中心一起举办的研讨会和研讨会

The proteomics Core D of the UAB Skin Disease Research Center provides support to pilot/feasibility awardees, SDRC-associated investigators and other skin disease researchers. This support consists of consultations imvolving two experienced senior faculty with expertise in 2D-gel proteomics and protein mass spectrometry, hands-on support by a junior faculty member skilled in several proteomics/mass spectrometry methods used and applied to skin disease research, training in the techniques offered by the Core, access to the instrumentation, and seminars by outside experts in proteomic and mass spectrometry. The specific services available in the Core include (1) development of experimental design and statistical considerations for experiments that include proteomics and mass spectrometry components; (2) 2D-gel separation of proteins from tissues, cells, subcellular fractions and specific body fluids (plasma/serum, urine, sweat, tears, tissue interstitial and blister fluids) - this involves isoelectric focusing/SDS-PAGE or clear native gel electrophoresis and makes use of Cy2, Cy3 and Cy5 fluorescent dyes and Sypro Ruby red post-staining, imaging and image analysis using DeCyder software, robotic gel spot picking, chemical processing and protease digestion, and peptide mass fingerprinting; (3) 1D-SDS-PAGE gel separation of membrane-associated proteins combined with nanoLC-ESI-tandem mass spectrometry; (4) isotope-labeling methods (iTRAQ reagents) to carry out quantitative analysis of the effects of treatments and time on members of the tissue, cell and fluid proteomes; and (5) development of specific multiple reaction ion monitoring (MRM) mass spectrometry methods to quantify peptides that allow quantitative analysis of individual, specified proteins. In addition, the LC-MRM-MS methods can be applied to the analysis of the reagents used in skin disease research, for example, green tea polyphenols and grape seed extract, as well as their and drug metabolites in the blood and other fluids. A special emphasis is made in maintaining quality control of the methods. A Laboratory Information Management System will be used to oversee the operations of the Core. Investigators will be trained individually in the use of the Core techniques as well as at workshops and a Symposium held in conjunction with the other NIH-funded Centers