Monoclonal Antibody and Associated Protein Facility

一个单克隆抗体核心设施是在该计划项目开始时创建的，用于生产推进该计划大部分研究所需的试剂。单克隆抗体是程序科学家常规使用的细胞和分子生物学方法的基础。自最初的资助期和前两次竞争性更新以来，单克隆抗体核心已经产生了许多单克隆抗体，这些单克隆抗体为该计划的成功做出了贡献。除了产生新的单克隆抗体外，核心还从公共领域收集了许多对程序科学家特别有用的杂交瘤。在这个更新申请中，我们建议扩大单克隆抗体核心成分B的范围，包括必要的设备和支持人员，以促进鉴定与项目成员正在研究的特定蛋白质物理相关的蛋白质伴侣。一段时间以来，对病毒蛋白进行免疫沉淀，然后用sds -聚丙烯酰胺凝胶分离，用考马斯银或胶体银染色已成为一种有用的鉴定相关病毒蛋白的实用技术

A monoclonal antibody core facility was created at the inception of this program project to produce reagents necessary for advancement of much of the research of the program. Monoclonal antibodies are fundamental to the cellular and molecular biologic approaches that the program scientists routinely use. Since the original funding period and the previous two competitive renewals, the monoclonal antibody core has generated many monoclonal antibodies that have contributed to the success of the program. In addition to the generation of novel monoclonal antibodies, the core has collected many hybridomas from the public domain that are particularly useful to the program scientists. In this renewal application, we propose to expand the scope of the monoclonal antibody core component B to include the necessary equipment and support staff to facilitate the identification of protein partners physically associated with specific proteins under study by program members. For some time now, immunoprecipitation for a viral protein followed by separation in an SDS-polyacrylamide gel and staining with Coomassie or colloidal silver has become a useful and practical technique for identification of associated proteins. In addition, use of epitope tagged versions of viral and cellular proteins enables the program members to identify associated proteins of practically any gene. Recent innovations and technical advances have made the large-scale identification of associated protein complexes feasible and practical. The increased availability and sensitivity of mass spectroscopy has reduced the amount of protein required for identification and the sequencing of the human and murine genomes now makes protein identification from partial sequences practical. As described in many of the specific projects, the use of this technology has facilitated progress in many areas, and it is clear that program scientists will benefit by having a dedicated resource that will facilitate preparation of suitable cells, preparative scale growth of cells, preparative scale immunoprecipitation, and sample preparation for mass spectroscopy.