| Catalase Activity Assay in Candida glabrata
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Author:
Date:
2014-03-20
[Abstract] Commensal and pathogenic fungi are exposed to hydrogen peroxide (H2O2) produced by macrophages of the host. Pathogenic fungi counteract the harmful effects of H2O2 with the enzyme catalase (EC 1.11.1.6), which decomposes two molecules of H2O2 to two molecules of H2O and O2. Contribution of antioxidant systems on fungal virulence is actively studied. Measurement of catalase activity can contribute to the elucidation of the factors that influence the regulation of this pivotal enzyme. Here we describe a simple spectrophotometric method in which the activity of catalase is measured in total yeast extracts. Decomposition of H2O2 by the yeast extract is followed by the decrease in ...
[摘要] 共生和致病真菌暴露于由宿主的巨噬细胞产生的过氧化氢（H 2 O 2 O 2）。 致病真菌抵消了H 2 O 2对于过氧化氢酶（EC 1.11.1.6）的有害影响，所述过氧化氢酶分解两个分子的H 2 O 2 - O 2至两个H 2 O和O 2分子。 积极研究抗氧化系统对真菌毒力的贡献。 过氧化氢酶活性的测量可有助于阐明影响这种关键酶的调节的因素。 在这里我们描述一个简单的分光光度法，其中过氧化氢酶的活性在总酵母提取物中测量。 通过酵母提取物的H 2 O 2 O 2分解后，在240nm处的吸光度降低。 吸光度随时间的差异（ΔA240）被推断为过氧化氢酶活性的量度。
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| Cellular Extract Preparation for Superoxide Dismutase (SOD) Activity Assay
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Author:
Date:
2013-07-05
[Abstract] Superoxide dismutase (SOD) acts as a primary defence against reactive oxygen species (ROS) by converting O2- to O2 and H2O2. Members of this enzyme family include CuZnSOD, MnSOD and FeSOD. Most eukaryotes harbor CuZnSOD and MnSOD, and FeSOD is found in plants and prokaryotes. This protocol is to demonstrate how to prepare the cellular extract for the identification and characterization of SODs in planta.
[摘要] 超氧化物歧化酶（SOD）通过将O 2转化为O 2和H 2 O而作为对活性氧（ROS）的主要防御 2 。 该酶家族的成员包括CuZnSOD，MnSOD和FeSOD。 大多数真核生物具有CuZnSOD和MnSOD，FeSOD存在于植物和原核生物中。 该协议是为了说明如何准备细胞提取物用于在植物中鉴定和表征SOD。
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| Flow Cytometric Detection of Reactive Oxygen Species
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Author:
Date:
2013-04-20
[Abstract] Reactive oxygen species (ROS) are molecules containing hydroxyl radicals or peroxides with unpaired electrons. In healthy aerobic cells, ROS are produced naturally as a byproduct of oxidative phosphorylation, oxidoreductase enzymes, or metal catalyzed oxidation at a controlled rate. However, ROS can be induced under some stress conditions especially exposure to environmental oxidants and certain drugs that leads to oxidative stress. Exceed ROS can cause damages in the building blocks of cells including DNA, proteins, and lipids, and eventually results in cell death. Cell-permeant 2', 7'-dichlorodihydrofluorescein diacetate (H2DCFDA) is a widely used ROS indicator. The reduced non-fluorescent fluorescein H2DCFDA can be oxidized and converted into fluorescent 2’, ...
[摘要] 活性氧（ROS）是含有羟基自由基的分子或具有不成对电子的过氧化物。 在健康好氧细胞中，ROS作为氧化磷酸化，氧化还原酶或金属催化氧化的副产物以受控速率天然产生。 然而，ROS可以在一些应激条件下诱导，特别是暴露于环境氧化剂和导致氧化应激的某些药物。 Exceed ROS可导致细胞构建块（包括DNA，蛋白质和脂质）的损伤，并最终导致细胞死亡。 细胞渗透剂2'，7'-二氯二氢荧光素二乙酸酯（H 2 DCFDA）是广泛使用的ROS指示剂。 还原的非荧光荧光素H 2 DCFDA可以被氧化并通过细胞内ROS转化为荧光2'，7'-二氯荧光素（DCF）。 在该协议中，我们应用H 2 DCFDA标记细胞内ROS并通过流式细胞术检测DCF强度。
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