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Ethylene glycol-bis

乙二醇 - 双(2-氨基乙醚)-N,N,N'',N''-四乙酸

公司名称: Sigma-Aldrich
产品编号: E3889
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Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae
Author:
Date:
2020-03-05
[Abstract]  Superoxide dismutases (SODs) act as a primary defence against reactive oxygen species (ROS) by converting superoxide anion radicals (O2-) into molecular oxygen (O2) and hydrogen peroxide (H2O2). Members of this enzyme family include CuZnSODs, MnSODs, FeSODs, and NiSODs, depending on the nature of the cofactor that is required for proper activity. Most eukaryotes, including yeast, possess CuZnSOD and MnSOD. This protocol aims at assessing the activity of the yeast Saccharomyces cerevisiae MnSOD Sod2p from cellular extracts using ... [摘要]  [摘要 ] 超氧化物歧化酶(SOD能)充当主防御针对反应性氧物质(ROS)通过转换的超氧阴离子自由基(O 2 - )为分子氧(O 2 )和过氧化氢(H 2 ? 2 )。这种酶的家庭成员包括CuZnSODs ,MnSODs ,FeSODs 和NiSODs ,这取决于是需要适当的活动辅助因子的性质。大多数真核生物,包括酵母,都具有CuZnSOD 和MnSOD 。该协议旨在评估酵母的活性 使用硝基蓝四唑染色法从细胞提取物中提取酿酒酵母MnSOD Sod2p 。该方法可用于估计Mn 2+ 的细胞生物利用度以及评估细胞的氧化还原状态。

[背景 ] 的SODs被定义为减少正常有氧代谢为氧气和过氧化氢期间形成的氧的有害自由基含金属的抗氧化剂酶。:这些酶是基于需要作为辅因子进行适当的酶活性的金属分类CuZnSODs ,MnSODs ,FeSODs ,和NiSODs 。在酿酒酵母中,有两个S OD :CuZn-Sod1p和Mn-Sod2p(Abreu和Cabelli ...

Isolation of Chromatin-bound Proteins from Subcellular Fractions for Biochemical Analysis
Author:
Date:
2018-10-05
[Abstract]  Shuttling of proteins between different cellular compartments controls their proteostasis and can contribute in some cases to regulate their activity. Biochemical analysis of chromatin-bound proteins, such as transcription factors, is often difficult because of their low yield and due to the interference from nucleic acids. This protocol describes a method to efficiently fractionate cells combined with a mechanical (i.e., sonication) or an enzymatic treatment (i.e., benzonase) that facilitates analysis of chromatin-bound protein extracts by Western blot analysis or by ... [摘要]  在不同细胞区室之间穿梭蛋白质控制它们的蛋白质稳态,并且在某些情况下可以有助于调节它们的活性。 染色质结合蛋白(例如转录因子)的生化分析通常是困难的,因为它们的产率低并且由于核酸的干扰。 该协议描述了一种有效分离细胞的方法,结合机械(即,超声处理)或酶处理(即,benzonase),有助于分析染色质结合蛋白提取物 通过蛋白质印迹分析或蛋白质下拉分析。 该方法对于富集特定亚细胞级分内的特定蛋白质以研究特定的翻译后修饰模式或鉴定特定的蛋白质 - 蛋白质相互作用可能是有价值的。
【背景】许多染色质结合蛋白的活性和翻译后调节研究很少,因为在分离它们进行生化分析时存在技术困难。这甚至是转录因子的情况,例如基本的螺旋 - 环 - 螺旋(bHLH)转录因子,其通常在组织或细胞模型中具有稀缺的时间和空间表达模式(Dennis 等。,2018)。当生物材料的量成为研究分子途径的障碍时,协议细化有助于解除技术限制(Gillotin和Guillemot,2016)。在我们最近的研究中,我们努力了解神经元bHLH转录因子Ascl1的蛋白水解是如何在神经元分化的细胞模型中调节的(Gillotin et ...

Metal-tagging Transmission Electron Microscopy for Localisation of Tombusvirus Replication Compartments in Yeast
Author:
Date:
2018-04-20
[Abstract]  Positive-stranded (+) RNA viruses are intracellular pathogens in humans, animals and plants. To build viral replicase complexes (VRCs) viruses manipulate lipid flows and reorganize subcellular membranes. Redesigned membranes concentrate viral and host factors and create an environment that facilitates the formation of VRCs within replication organelles. Therefore, efficient virus replication depends on the assembly of specialized membranes where viral macromolecular complexes are turned on and hold a variety of functions. Detailed characterization of viral replication platforms in cells ... [摘要]  正链(+)RNA病毒是人,动物和植物中的细胞内病原体。构建病毒复制酶复合物(VRC)病毒操纵脂质流动和重组亚细胞膜。重新设计的膜集中了病毒和宿主因子,并创造了促进复制细胞器内VRC形成的环境。因此,有效的病毒复制取决于病毒大分子复合物开启并具有各种功能的特殊膜的组装。细胞中病毒复制平台的详细特征需要复杂的成像方法。在这里我们提出一个协议,用肉眼标记透射电子显微镜(METTEM)可视化酵母中的tombusvirus复制酶复合物的三维组织。该协议使我们能够用METTEM和电子断层扫描成像三维病毒复制酶分子的细胞内分布。我们的研究显示病毒复制酶分子如何在特化细胞膜内构建复制复合物。

【背景】正链RNA病毒的复制取决于细胞膜的重塑。细胞内膜作为VRC装配的结构支架,提供调节病毒复制酶活性和保护病毒RNA免受宿主抗病毒防御的必需脂质和辅因子(Miller和Krijnse-Locker,2008; den Boon <等,2010; nagy和pogany,2011;="" nagy,2016)。电子显微镜观察到具有活性vrc的复制细胞器的结构。="" vrc以单个膜囊或'小球',管状球形立方体膜,双膜囊泡(dmv)或平面寡聚体阵列装配(de="" castro等人,2013)。通常在rna病毒感染的细胞中观察到小球。它们通过在各种细胞器中内陷而形成,并具有对胞质溶胶的狭窄开口(den="" boon="" et=""> ...

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