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Monoclonal Antibody Resources for C. elegans

线虫单克隆抗体资源

基本信息

批准号：
7250855
负责人：
MICHAEL L NONET
金额：
$ 28.82万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-30 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7250855
关键词：
Affinity Animal Model Antibodies Antigen Targeting Antigens Ascites Biochemical Biology Caenorhabditis elegans Caenorhabditis elegans Proteins Cell Fractionation Cell Line Cells Centrosome Communities Complex Cultured Cells Data Detection Development Developmental Cell Biology Drosophila inturned protein Dynein ATPase Embryo Endoplasmic Reticulum Endosomes Epitopes Golgi Apparatus Green Fluorescent Proteins Hybridomas Immunoprecipitation In Situ Individual Intermediate Filaments Iowa Kinesin Label Laboratory Research Localized Macromolecular Complexes Marketing Membrane Methodology Molecular Monoclonal Antibodies Nervous system structure Nuclear Envelope Numbers Organelles Oryctolagus cuniculus Phosphorylation Post-Translational Protein Processing Process Proteins Reagent Replication Origin Research Research Personnel Resources Series Serum Specificity Subcellular structure Synaptic Vesicles Testing Time Tube Vertebrates Vesicle Western Blotting Work cost immunocytochemistry in vivo multicatalytic endopeptidase complex mutant polyclonal antibody programs protein protein interaction research study tool

项目摘要

Antibodies (Abs) are widely used to detect specific cellular components both in situ and in biochemical experiments aimed at defining protein-protein interactions, protein processing, and reversible protein modifications such as phosphorylation. Polyclonal Abs are relatively simple to make and are widely used in molecular and cellular studies. However, such reagents are limited in quantity and thus rarely made widely available. Monoclonal Abs are more complicated to make, but hybridomas secreting the monoclonal antibody can be passaged ad infinitum permitting the identical reagent to be used by many different research group over long periods of time. Polyclonal and monoclonal Abs are available commercially that are directed against a huge array of vertebrate cellular components. By contrast, few such commercial reagents are available for work with model organisms such as C. elegans probably because the market for these reagents is too small to make them commercially viable. I propose to develop a 'tool kit'consisting of monoclonal Abs directed against a series of proteins that label discrete cellular components and subcellular compartments of C. elegans. The compartments include organelles common to all cells (e.g. endoplasmic reticulum, the Golgi, endosomes), organelles of specialized cells (e.g.synaptic vesicles), subcellular structures common to all cells (e.g.nuclear membrane, centrosomes and caveoli) and components that label domains in the developing embryo, germline, and nervous system. The cellular components include macromolecular complexes (e.g.splicesomes, proteosomes, replication origin complexes) and common cellular proteins (e.gclathrin, kinesin, dynein and intermediate filaments). The Abs will be thoroughly characterized and made publically available through the Developmental Studies Hybridoma Bank. The tool kit will be extremely useful for C. elegans researchers. It will provide a set of standardized renewable reference antibody tools available to the entire community. Secondly, it will greatly facilitate localizing other cellular components by providing a large set of cellular markers that can be used coordinately with rabbit polyclonal antibodies for co-localization studies. Finally, it will stimulate the use of biochemical approaches, such as subcellular fractionation and immunoprecipitations, by providing means to detect specific cellular compartments in extracts and the presence of specific components in immunoprecipitates.

抗体（Abs）被广泛用于原位和生化检测特定的细胞成分。旨在定义蛋白质-蛋白质相互作用，蛋白质加工和可逆蛋白质的实验修饰如磷酸化。多克隆抗体制备相对简单，并且广泛用于分子和细胞研究。然而，这些试剂的数量有限，因此很少广泛生产 available.单克隆抗体的制备更为复杂，但分泌单克隆抗体的杂交瘤抗体可以无限传代，从而允许相同的试剂用于许多不同的研究。集团在很长一段时间。多克隆和单克隆Ab可商购获得，其针对对抗大量脊椎动物细胞成分相比之下，很少有这样的商业试剂是可用于与模式生物如C.可能是因为市场上试剂太小，无法使其在商业上可行。我建议开发一个"工具包"，针对一系列蛋白质的单克隆抗体，这些蛋白质标记离散的细胞组分和亚细胞组分， C.优雅的隔室包括所有细胞共有的细胞器（例如，内质网）。网状细胞、高尔基体、核内体）、特化细胞的细胞器（例如突触囊泡）、亚细胞所有细胞共有的结构（如核膜、中心体和小窝）和标记发育中的胚胎、生殖系和神经系统中的结构域。蜂窝组件包括大分子复合物（例如剪接体、蛋白体、复制起点复合物）和常见的细胞蛋白质（例如网格蛋白、驱动蛋白、动力蛋白和中间丝）。Abs将彻底通过发展研究杂交库进行表征并提供。工具试剂盒对C. elegans研究人员。它将提供一套标准化的可再生能源提供给整个社区的参考抗体工具。其次，它将极大地促进其他通过提供大量的细胞标记物，用于共定位研究的多克隆抗体。最后，它将促进生物化学方法的使用，例如亚细胞分级分离和免疫沉淀，提取物中的隔室和免疫沉淀物中特定组分的存在。