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D-(+)-Glucose

D-(+)-葡萄糖

Company: Sigma-Aldrich
Catalog#: G8270
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Preparation and Purification of Proteins Secreted from Phytophthora sojae
Author:
Date:
2018-10-20
[Abstract]  Phytophthora sojae, the causal agent of soybean root and stem rot, is responsible for enormous economic losses in soybean production. P. sojae secrets various effectors to reprogram host immunity. The plant apoplastic space is a major battleground in plant-pathogen interactions. Here we describe a protocol for purification and isolation of secreted proteins from P. sojae, including precipitation of secreted proteins from P. sojae culture filtrate, chromatographic purification of the secreted proteins and analysis of the proteins by Mass spectrometry. With this protocol, it will be easier to identify potential apoplastic effectors in Phytophthora and will benefit our understanding of plant-microbe interactions. [摘要]  大豆根和茎腐病的致病因子大豆疫霉(Phytophthora sojae)是造成大豆生产的巨大经济损失的原因。 P上。 sojae >秘密各种效应器重新编程宿主免疫力。植物质外体空间是植物 - 病原体相互作用的主要战场。在这里,我们描述了从 P中纯化和分离分泌蛋白的方案。 sojae >,包括来自 P的分泌蛋白的沉淀。 sojae >培养滤液,分泌蛋白的色谱纯化和质谱分析蛋白质。通过该协议,可以更容易地鉴定 Phytophthora >中潜在的质外体效应物,并有助于我们对植物 - 微生物相互作用的理解。

【背景】从 Phytophthora >物种中分泌的蛋白质的纯化对于理解 Phytophthora >发病机理至关重要。过去,V8果汁和植物(番茄和利马豆)果汁培养基已用于培养 Phytophthora >,培养物过滤后用于分析 Phytophthora >分泌的蛋白质。这些方案的缺点是培养基含有大量的植物蛋白,它们代表了大部分检测到的蛋白质。在该方案中,我们使用了不含任何蛋白质的合成液体培养基。该培养基显着降低了 Phytophthora >培养滤液的背景。此外,利用凝胶过滤脱盐和筛分柱代替离子交换柱,可以有效和大规模纯化 Phytophthora >分泌的蛋白质。

Measurement of Dopamine Using Fast Scan Cyclic Voltammetry in Rodent Brain Slices
Author:
Date:
2018-10-05
[Abstract]  Fast scan cyclic voltammetry (FSCV) is an electrochemical technique that allows sub-second detection of oxidizable chemical species, including monoamine neurotransmitters such as dopamine, norepinephrine, and serotonin. This technique has been used to record the physiological dynamics of these neurotransmitters in brain tissue, including their rates of release and reuptake as well as the activity of neuromodulators that regulate such processes. This protocol will focus on the use of ex vivo FSCV for the detection of dopamine within the nucleus accumbens in slices obtained from rodents. We have included all necessary materials, reagents, recipes, procedures, and analyses in order to successfully perform this technique in the laboratory setting. Additionally, we have also included ... [摘要]  快速扫描循环伏安法(FSCV)是一种电化学技术,可以亚秒检测可氧化的化学物质,包括单胺类神经递质,如多巴胺,去甲肾上腺素和血清素。 该技术已被用于记录这些神经递质在脑组织中的生理动态,包括它们的释放和再摄取速率以及调节这些过程的神经调节剂的活性。 该方案将集中于使用离体 FSCV检测从啮齿动物获得的切片中的伏隔核内的多巴胺。 我们已经包含了所有必要的材料,试剂,配方,程序和分析,以便在实验室环境中成功地执行该技术。 此外,我们还提供了一些警示点,我们认为这些警告点对于那些在该领域的新手有帮助。

【背景】由于首先将用于检查生理系统的电特性的能力用于临床前科学研究,因此已经开发了许多用于研究突触生理学的技术。从鱿鱼轴突电生理记录的早期到现在的快速扫描循环伏安法(FSCV)在人类帕金森病患者中进行(Kishida et al。,2016; Lohrenz et al。,2016),该领域在相对较短的时间内取得了重大进展。该协议的重点是FSCV,是物理学家,分析化学家和神经科学家之间40多年的创新和合作的技术成果。早在19世纪(Bard和Zoski,2000年),电化学与Michael Faraday和Alessandro Volta一起诞生(Bard and Zoski,2000),直到20世纪20年代,Jaroslav ...

High-throughput Microscopic Analysis of Salmonella Invasion of Host Cells
Author:
Date:
2018-09-20
[Abstract]  Salmonella is a Gram-negative bacterium causing a gastro-enteric disease called salmonellosis. During the first phase of infection, Salmonella uses its flagella to swim near the surface of the epithelial cells and to target specific site of infection. In order to study the selection criteria that determine which host cells are targeted by the pathogen, and to analyze the relation between infecting Salmonella (i.e., cooperation or competition), we have established a high-throughput microscopic assay of HeLa cells sequentially infected with fluorescent bacteria. Using an automated pipeline of image analysis, we quantitatively characterized a multitude of parameters of infected and non-infected cells. Based on this, we established a predictive model that ... [摘要]  沙门氏菌是革兰氏阴性细菌,引起称为沙门氏菌病的胃肠疾病。在感染的第一阶段,沙门氏菌使用其鞭毛在上皮细胞表面附近游泳并靶向特定的感染部位。为了研究确定哪种宿主细胞被病原体靶向的选择标准,并分析感染沙门氏菌( ie ,合作或竞争)之间的关系,我们有建立了对荧光细菌依次感染的HeLa细胞的高通量显微镜检测。使用自动化图像分析管道,我们定量表征了感染和未感染细胞的众多参数。基于此,我们建立了一个预测模型,使我们能够识别宿主细胞易受感染的参数。我们发现宿主细胞易损性有两个来源:病原体诱导的细胞易感性从沙门氏菌摄取中出现并持续存在于感染过程的后期阶段;以及与细胞固有属性相关的宿主细胞固有的脆弱性,例如局部细胞拥挤和胆固醇含量。我们的方法基于形态学或分子宿主细胞参数预测单层上皮细胞中沙门氏菌感染的概率。在这里,我们提供了工作流程的详细描述,包括基于计算机的分析管道。我们的方法有可能应用于研究宿主 - 病原体相互作用的其他组合。

【背景】鼠伤寒沙门氏菌血清型鼠伤寒沙门氏菌通过摄入受污染的食物或水感染宿主,引起沙门氏菌病。一旦细菌到达肠道的远端回肠,它们就会侵入广泛的宿主细胞,包括肠上皮细胞(Watson和Holden,2010)。在宿主细胞入侵的第一阶段,沙门氏菌选择其目标,使用其鞭毛游泳并扫描上皮表面(Misselwitz et ...

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