CORE--ENZYMES AND ANTIBODIES FACILITY

酶和抗体设施核心的总体目的 是为酶的特性提供专业知识和设施 它们在环境化学物质的生物转化中非常重要。这个 应用程序提供将由提供的特定服务的详细列表 酶和抗体设施的核心。 酶和抗体中心成立于1996年10月，由 结合两个预先存在的核心，即单抗生产 和化验实验室(由P.E.Thomas领导)和实验室 用于蛋白质的纯化和鉴定(由C.S. 杨)。新的核心设施由托马斯博士领导，它提供 连续性。托马斯博士将由洪博士协助，他将监督 核心设施-S在为表达提供专业知识和资源方面的新角色 重组酶，用于分析猪的遗传多态 异源生物代谢酶(XME、S)和DNA探针的制备。 酶和抗体设施的核心设在三个实验室中 不同的建筑：EOSHI大楼，癌症研究实验室 (位于EOSHI大楼旁边)和RWJMS的Chada？S博士实验室 (也位于EOSHI大楼旁边)。这一核心设施很容易 中心成员均可访问。 酶和抗体设备核心公司生产出了最好的，如果不是 BEST，各种P450的单抗和多克隆抗体清单 酶和其他异源代谢酶(XME)。库存是 令人印象深刻的不仅是抗体的数量 准备好了，但也是为了这些抗体已经达到的程度 其特征在于它们的特殊性和特性(例如 他们是否识别西方免疫印迹上变性的P450酶，以及 无论它们是否抑制P450依赖的代谢，这是一种非常可取的特性 这种抗体)。酶和抗体设施核心支持一种 中心开展的研究项目数量和种类令人印象深刻 会员。该装置用于制备重组P450酶 甲基叔丁基醚(MTBE)代谢的研究 Hong)；纯化一氧化氮合酶的诱导型P450酶和 芒果醛交联蛋白(P.E.Thomas、J.Laskin和G.Witz)； 制备抗血红素加氧酶的单抗(针对伊巴分枝杆菌)； 提供用于免疫定量和免疫细胞化学的抗体(对于M.Iba， 首页--期刊主要分类--期刊细介绍--期刊题录与文摘--期刊详细文摘内容 众多外部调查人员)；进行抗体抑制实验以 确定单个P450酶对MTBE代谢的贡献 (对于杨振星和洪振英)和雌二醇(对于B.T.朱和A.H.康尼)； 并筛查人类编码细胞色素P450 2A6、细胞色素P450 2 E1、 Pi和Mu形成谷胱甘肽S转移酶和苯醌氧化还原酶(For J.Y.Hong和S.Mohr)。酶和抗体设施核心还提供 接受各种免疫化学和分子生物学技术方面的培训。 酶和抗体设施核心由成员在每个 研究核心(尽管程度不一)，以及许多外部 研究人员。 该应用程序确定了大量未来的研究项目， 将得到酶和抗体设施核心的支持。

The overall purpose of the Enzymes and Antibodies Facility Core is to provide expertise and facilities for the characterization of enzymes that are important in the biotransformation of environmental chemicals. The application provides a detailed list of specific services to be provided by the Enzymes and Antibodies Facility Core. The Enzyme and Antibody Facility Core was formed in October of 1996 by combining two pre-existing Cores, namely the Monoclonal Antibody Production and Assay Laboratory (which was directed by P.E. Thomas), and the Laboratory for Protein Purification and Characterization (which was directed by C.S. Yang). The new Core Facility is directed by Dr. Thomas, which provides continuity. Dr. Thomas will be assisted by Dr. Hong, who will oversee the Core Facility?s new role in providing expertise and resources for expressing recombinant enzymes, for analyzing genetic polymorphisms in xenobiotic-metabolizing enzymes (XME?s) and for preparing DNA probes. The Enzyme and Antibody Facility Core is located in laboratories in three different buildings: the EOSHI building, the Laboratory for Cancer Research (located next to the EOSHI building) and Dr. Chada?s laboratory in the RWJMS (also located next to the EOSHI building). This Core Facility is readily accessible to members of the Center. The Enzyme and Antibody Facility Core has produced one of best, if not the best, inventory of monoclonal and polyclonal antibodies against various P450 enzymes and other xenobiotic-metabolizing enzymes (XME). The inventory is impressive not only in terms of the number of antibodies that have been prepared, but also for the degree to which these antibodies have been characterized with respect to their specificity and properties (such as whether they recognize the denatured P450 enzyme on western immunoblots, and whether they inhibit P450-dependent metabolism, a highly desirable property of such antibodies). The Enzyme and Antibody Facility Core has supported an impressive number and variety of research projects carried out by Center members. The facility was used to prepare recombinant P450 enzymes for studies of methyl-tert-butyl ether (MTBE) metabolism (for C.S. Yang and J.Y. Hong); to purify P450 enzymes, the inducible form of nitric oxide synthase and muconaldehyde cross-linked proteins (for P.E. Thomas, J. Laskin and G. Witz); to prepare monoclonal antibodies against heme oxygenase (for M. Iba); to provide antibodies for immunoquantitation and immunocytochemistry (for M. Iba, D. Morse, H. Lowndres, C.S. Yang, J. Landau, C. Gardner, D. Laskin and numerous outside investigators); to conduct antibody-inhibition experiments to identify the contribution of individual P450 enzymes to the metabolism of MTBE (for C.S. Yang and J.Y. Hong) and estradiol (for B.T. Zhu and A.H. Conney); and to screen for polymorphisms in the human genes encoding CYP2A6, CYP2E1, the pi and mu forms glutathione S-transferases and quinone oxidoreductase (for J.Y. Hong and S. Mohr). The Enzyme and Antibody Facility Core also provides training in a variety of immunochemical and molecular biology techniques. The Enzyme and Antibody Facility Core is used by members in each of the research cores (albeit to varying extent), and by numerous outside researchers. The application identifies a large number of future research projects that will be supported by the Enzyme and Antibody Facility Core.