CORE--ANTIOXIDANT ENZYME

说明(申请人提供)：抗氧化酶的用途 核心(AEC)是提供抗氧化酶的专业知识和分析 个人研究项目。项目调查员和项目调查员之间的讨论 AEC主任将负责确定哪些实验和/或分析 跑。AEC工作人员将执行拟议的工作或向 关于如何进行实验的调查员。其中的一些工作， 特别是新的cDNA或在新的细胞系中的转染以及 反义试剂的发展，本质上是发展的。 AEC将协助西部、北部和南部的酶活性分析 分析。酶检测将在抗氧化酶铜和铜上进行 含锰的锌超氧化物歧化酶(CuZnSOD) 超氧化物歧化酶(MnSOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPX， 胞液和磷脂)。对于项目1，还将完成以下工作 白细胞酶髓过氧化物酶(MPO)。对于试剂，我们有抗体 以及所有主要抗氧化物酶的cDNA探针。此外，AEC将 帮助将这些酶的cDNA导入感兴趣的细胞系 项目调查员。我们有几种方法的经验 以及针对这些酶的许多不同的表达载体。 在这项工作中，AEC将继续保持和发展 抗氧化剂的试剂，即抗体、cDNA和表达载体 蛋白质。AEC还将发展反义药物作为目标 抑制所有主要抗氧化剂蛋白质水平的试剂。

DESCRIPTION (provided by applicant): The purpose of the Antioxidant Enzyme Core (AEC) is to provide antioxidant enzyme expertise and analysis for the individual research projects. Discussion between a project investigator and the AEC Director will be held to determine which experiments and/or assays are run. The AEC staff will carry out the proposed work or will advise the investigator on how to perform the experiments. Some of this work, particularly transfections of new cDNAs or in new cell lines as well as the development of antisense reagents, will be developmental in nature. The AEC will aid in enzyme activity assays, western, northern, and Southern analysis. The enzyme assays will be run on the antioxidant enzymes copper-and zinc-containing superoxide dismutase (CuZnSOD), manganese-containing superoxide dismutase (MnSOD), catalase (CAT), and glutathione peroxidase (GPx, both cytosolic and phospholipid). For Project 1, work will also be done on the white cell enzyme myeloperoxidase (MPO). For reagents, we have antibodies and cDNA probes to all primary antioxidant enzymes. In addition, the AEC will aid in transfecting the cDNAs for these enzymes into cell lines of interest to the project investigators. We have experience with several methods of transfection, as well as many different expression vectors for these enzymes. In the course of this work, the AEC will continue to maintain and develop reagents i.e., antibodies, cDNAs, and expression vectors for the antioxidant proteins. The AEC will also have as a goal the development of antisense reagents to inhibit the levels of all of the major antioxidant proteins.