Skin Proteomics Core

皮肤蛋白质组学核心

基本信息

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
项目类别：
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

项目摘要

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为飞行员/可行性获奖者提供了支持， SDRC相关的研究人员和其他皮肤病研究人员。此支持包括咨询在2D-GEL蛋白质组学和蛋白质质谱学方面拥有两位经验丰富的高级教师，初级教师的动手支持，熟练于几种蛋白质组学/质谱方法并应用于皮肤疾病研究，核心提供的技术培训，进入仪器和蛋白质组学外部专家的研讨会。特定服务在核心中可用，包括（1）开发实验设计和统计注意事项包括蛋白质组学和质谱分量的实验；（2）蛋白质的2凝结分离来自组织，细胞，亚细胞级分和特定的体液（血浆/血清，尿液，汗水，泪水，撕裂，组织间质和水泡流体） - 这涉及等电聚焦/SDS -PAGE或清除天然凝胶电泳并利用CY2，CY3和CY5荧光染料以及Sypro红宝石红色后染色，成像和图像使用Decyder软件，机器人凝胶点采摘，化学加工和蛋白酶消化以及肽质量指纹；（3）膜相关蛋白与结合的膜相关蛋白的1D-SD-PAGE凝胶分离 Nanolc-Esi-Tandem质谱；（4）同位素标记的方法（ITRAQ试剂）执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；（5）开发特定的多重反应离子监测（MRM）质谱法量化允许对个体，指定蛋白质进行定量分析的肽。另外，LC-MRM-MS 方法可以应用于对皮肤病研究中使用的试剂的分析，例如绿茶多酚和葡萄种子提取物及其血液和其他液体中的药物代谢物。特别重点是维持对方法的质量控制。实验室信息管理系统将用于监督核心的操作。调查人员将对核心的使用进行单独培训技术和研讨会和与其他NIH资助的中心共同举行的研讨会