CORE--ANTIGEN AND ANTIBODY PRODUCTION

说明(由申请人提供)：抗原的主要用途和 抗体生产共享资源是提供及时、经济的 GST融合蛋白、小鼠多克隆抗体和单抗， 可以为圣安东尼奥癌症研究所的成员生产腹水。 高质量抗体的可获得性是 了解基因功能以及在随后的翻译应用中的应用。 SACI成员识别或研究对癌症重要的新基因 易感、进展和预后。GST融合蛋白更具优势 用于生产和筛选自融合以来的高质量抗体 通过简单的操作可以提纯蛋白质。最纯净的GST-融合 蛋白质能刺激小鼠的良好免疫反应。虽然GST-Fusion 蛋白质是核心最常用的抗原，其他方法可以 如有必要或调查员愿意，可使用。核心是定向的 由芭芭拉·A·克里斯蒂博士撰写，她在制作 多种多克隆和单克隆抗体。她的实验室创造了， 纯化和利用多个GST-融合蛋白和His-标记蛋白 并在杆状病毒中表达了几种蛋白。这些蛋白质有 用于多克隆抗体的生产，单抗的生产， 抗体亲和纯化、DNA结合研究、蛋白质-蛋白质 蛋白质相互作用研究、表达文库筛选及作为底物的研究 激活酶分析。此外，克里斯蒂博士的实验室在使用 这些抗体用于分析细胞内蛋白质的表达和功能 哺乳动物细胞，免疫荧光，免疫组织化学，免疫沉淀 和亚细胞分馏。 该共享资源提供的特定服务包括GST-融合蛋白 制备、小鼠多克隆抗血清制备、小鼠单抗 生产，腹水生产，使用这些试剂来表征 特异性基因产物及杂交瘤细胞株和抗体的冷冻保存 股票。

DESCRIPTION (provided by applicant): The primary purpose of the Antigen and Antibody Production Shared Resource is to provide a timely and economical means by which GST-fusion protein, mouse polyclonal and monoclonal antibodies, and ascites can be produced for members of the San Antonio Cancer Institute. Availability of high quality antibodies represents a critical element for the understanding of gene function and in subsequent translational applications. SACI members identify or work on new genes that are important for cancer predisposition, progression and prognosis. GST fusion proteins are superior for the production and selection of high quality antibodies since the fusion protein can be purified through simple manipulation. Most purified GST-fusion proteins stimulate excellent immunoresponses in mice. Although GST-fusion proteins are most commonly used as antigens by the core, other approaches can be used if necessary or preferred by the investigator. The core is directed by Barbara A. Christy, Ph.D., who has experience in the production of a variety of polyclonal and monoclonal antibodies. Her laboratory has created, purified and used a number of GST-fusion proteins and His-tagged proteins in bacteria and expressed several proteins in baculovirus. These proteins have been used for polyclonal antibody production, monoclonal antibody production, antibody affinity purification, DNA binding studies, protein-protein interaction studies, expression library screening and as substrates in protein kinase assays. In addition, Dr. Christy's lab is experienced in the use of these antibodies for the analysis of protein expression and function in mammalian cells, immunofluorescence, immunohistochemistry, immunoprecipitation and subcellular fractionation. Specific services provided by this Shared Resource include GST-fusion protein production, mouse polyclonal antisera production, mouse monoclonal antibody production, ascites production, the use of these reagents in characterizing specific gene products, and cryopreservation of hybridoma lines and antibody stocks.