权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

CORE C: PROTEIN EXPRESSION AND CELL MORPHOLOGY

核心 C：蛋白质表达和细胞形态

基本信息

批准号：
8168465
负责人：
Sean M Ward
金额：
$ 23.18万
依托单位：
UNIVERSITY OF NEVADA RENO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8168465
关键词：
3-Dimensional Antibodies Area Binding Sites Cell Culture Techniques Cells Cellular Morphology Complex Computer Retrieval of Information on Scientific Projects Database Confocal Microscopy Cryoelectron Microscopy Cultured Cells Data Detection Disease Electron Microscopy Fluorescence Funding Grant Hypertrophy Image Imagery Immunofluorescence Immunologic Immunoprecipitation Individual Institution Label Laboratories Measurement Microfilaments Mus Muscle Myosin Light Chain Kinase Negative Staining Nevada Nomarski Interference Contrast Microscopy Non-Insulin-Dependent Diabetes Mellitus Phase Phenotype Phosphoric Monoester Hydrolases Population Heterogeneity Preparation Protein Overexpression Proteins Proteomics Protocols documentation Research Research Personnel Resources Sampling Services Site Smooth Muscle Myocytes Sorting - Cell Movement Source Structure Support System Tissues Transfection United States National Institutes of Health Visceral Western Blotting digital gel electrophoresis light microscopy protein expression reconstruction sample fixation tissue preparation

项目摘要

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. service for the examination of protein expression using western analysis combined with a variety of morphological approaches to localize levels of expression in tissues with heterogeneous populations of cells. The Core has integrated western analysis and morphological studies such that quantitative protein expression and imaging needs from basic morphological to complex 3-dimensional volumetric measurements or electron microscopy will be performed in the same laboratory providing a seamless support mechanism for individual projects. The services to be provided by the Core include protein overexpression, protein expression analysis in tissues and cells, transfections of cells, cell culture, cell lysate preparations, immunoprecipitations, tissue preparations, western blots, gel electrophoresis, nanoLC and MALDl sample preparation. The Core will also act as an interface with the Nevada Proteomics Core for mass spectral analysis and analysis of MALDl data. Integrated within this support system with be a structural analysis of tissues and visualization of the sites of protein expression within tissues and isolated cells. Support will be provided in the areas of light microscopy (bright field, phase contrast and differential interference contrast microscopy) for the proposed histological studies on visceral tissues. The Core will also provide morphological expertise in the areas of immunofluorescence (regular fluorescence and confocal microscopy) for the examination of protein expression in heterogeneous tissues that contain several cellular phenotypes. The Core will troubleshoot the use of antibodies (i.e. fixation protocols, non-specific labeling) to optimize use and the detection of proteins. Digital reconstructions will be deconvolved and 3-dimensional volume reconstructions performed when needed The core will also provide support for the examination of eGFP expression in tissues and isolated cells prior to the sorting of smooth muscle cells from smMHCICreleGFP mice. The Core will also provide expertise in the ultrastructural analysis of muscles and isolated myofilaments. Several areas of expertise will be supported including conventional electron microscopy, negative staining of isolated myofilaments, cryoelectron microscopy and immunogold labeling of proteins. Ultrastructural analysis is necessary to determine the pathological changes that occur in smooth muscle cells during hypertrophy and changes in structure due to diseases like type II DM. Analysis of actin filaments using negative staining and immunogold labeling will be provided to determine the binding sites of myosin light chain kinase and phosphatase.

这个子项目是众多研究子项目之一