Skin Proteomics Core

皮肤蛋白质组学核心

• 批准号: 7677162
• 负责人: STEPHEN BARNES
• 金额: $ 12.17万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7677162
• 关键词: Animal Model; Arts; Biological; Blood; Body Fluids; Bulla; Cancer Center; Cells; Chemicals; Computer software; Consultations; Data; Databases; Dermatology; Detection; Development; Digestion; Disease; Educational Activities; Educational workshop; Epigallocatechin Gallate; Event; Experimental Designs; Faculty; Fingerprint; Fluorescent Dyes; Funding; Gel; Hand; Human Resources; Image; Image Analysis; Immunoprecipitation; Individual; Ions; Isoelectric Focusing; Isotope Labeling; Laboratories; Lead; Liquid substance; Management Information Systems; Mass Spectrum Analysis; Membrane Proteins; Methods; Modification; Monitor; Operative Surgical Procedures; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Plasma; Post-Translational Protein Processing; Preparation; Prevention; Principal Investigator; Process; Protein Analysis; Proteins; Proteome; Proteomics; Protocols documentation; Quality Control; Reaction; Reagent; Research; Research Personnel; Research Training; Resources; Robotics; Role; Sampling; Science; Services; Skin; Skin Tissue; Specific qualifier value; Spottings; Staining method; Stains; Subcellular Fractions; Sweat; Sweating; Techniques; Technology; Time; Tissues; Training; University of Alabama at Birmingham Cancer Center; Update; Urine; base; cyanine dye 5; experience; gel electrophoresis; grape seed extract; improved; instrumentation; interstitial; member; programs; protein complex; research study; skin disorder; small molecule; symposium; tandem mass spectrometry; treatment effect

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

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。这种支持包括咨询 聘请两位经验丰富、在 2D 凝胶蛋白质组学和蛋白质质谱分析方面具有专业知识的高级教师， 由精通多种蛋白质组学/质谱方法的初级教员提供实践支持 并应用于皮肤病研究、核心提供的技术培训、获得 仪器，以及由外部专家在蛋白质组学和质谱学方面举办的研讨会。具体服务内容 核心中提供的内容包括（1）开发实验设计和统计考虑因素 包括蛋白质组学和质谱分析组件的实验； (2) 蛋白质的二维凝胶分离 来自组织、细胞、亚细胞部分和特定体液（血浆/血清、尿液、汗液、眼泪、组织 间质和泡状液体） - 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 荧光染料以及 Sypro Ruby red 后染色、成像和图像 使用 DeCyder 软件、机器人凝胶点挑选、化学处理和蛋白酶消化进行分析，以及 肽质量指纹分析； (3) 1D-SDS-PAGE凝胶分离膜相关蛋白结合 nanoLC-ESI-串联质谱法； (4)同位素标记方法(iTRAQ试剂)进行 定量分析治疗和时间对组织、细胞和液体蛋白质组成员的影响； (5) 开发特定的多反应离子监测 (MRM) 质谱方法 对肽进行定量，从而可以对单个特定蛋白质进行定量分析。此外，LC-MRM-MS 该方法可应用于皮肤病研究中使用的试剂的分析，例如绿茶 多酚和葡萄籽提取物，以及它们和血液和其他体液中的药物代谢物。一个特别的 重点是保持方法的质量控制。实验室信息管理系统 将用于监督核心的运营。研究人员将接受核心使用方面的单独培训 技术以及与其他 NIH 资助的中心联合举办的研讨会和研讨会