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Sodium dodecyl sulfate

十二烷基硫酸钠

Company: Sigma-Aldrich
Catalog#: L3771
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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。

Detection of Cell Death in Planarians
Author:
Date:
2018-10-05
[Abstract]  Planarians are freshwater flatworms, well known for their ability to regenerate a complete organism from any piece of their body. Furthermore, planarians are constantly growing and degrowing throughout their lives, maintaining a functional and proportioned body. These properties rely on the presence of a population of adult stem cells and on the tight control of their cell renewal, which is based on the balance between the proliferation of new cells and their differentiation, and the death of unnecessary cells. Due to the importance of these two processes in planarian biology, over the years, researchers have optimized molecular techniques to detect both cell proliferation and cell death in planarians. Here, we present the two main protocols currently used for cell death detection and ... [摘要]  涡虫是淡水扁虫,因其能够从身体的任何部分再生完整的有机体而闻名。 此外,涡虫在其一生中不断生长和去除,保持功能和比例的身体。 这些特性依赖于成体干细胞群的存在以及对细胞更新的严格控制,其基于新细胞增殖与其分化之间的平衡以及不必要细胞的死亡。 由于这两个过程在涡虫生物学中的重要性,多年来,研究人员已经优化了分子技术,以检测涡虫中的细胞增殖和细胞死亡。 在这里,我们提出了目前用于细胞死亡检测和量化的两种主要方案:Caspase-3活性定量和TUNEL分析。
【背景】成体生物体中的细胞更新是基于三个过程的复杂机制:(a)通过细胞死亡消除选定的细胞; (b)通过细胞分裂取代已消除的细胞,通常涉及成体干细胞及其后代; (c)新生细胞的分化及其与先前存在的组织的整合(Pellettieri和Sanchez ...

Extraction and Quantification of Polyphosphate (polyP) from Gram-negative Bacteria
Author:
Date:
2018-09-20
[Abstract]  Polyphosphate (polyP), a universally conserved biomolecule, is composed of up to 1,000 phosphate monomers linked via phosphoanhydride bonds. Reaching levels in bacteria that are in the high nmoles per mg protein range, polyP plays important roles in biofilm formation and colonization, general stress protection and virulence. Various protocols for the detection of polyP in bacteria have been reported. These methods primarily differ in the ways that polyP is extracted and/or detected. Here, we report an improved method, in which we combine polyP extraction via binding to glassmilk with a very sensitive PolyP kinase/luciferase-based detection system. By using this procedure, we significantly enhanced the sensitivity of polyP detection, making it potentially applicable for mammalian tissues. [摘要]  多磷酸盐(polyP)是一种普遍保守的生物分子,由多达1,000个通过磷酸酐键连接的磷酸盐单体组成。 达到每毫克蛋白质高纳摩尔细菌的水平,polyP在生物膜形成和定植,一般应力保护和毒力中起重要作用。 已经报道了用于检测细菌中polyP的各种方案。 这些方法主要在于提取和/或检测polyP的方式不同。 在这里,我们报告了一种改进的方法,其中我们结合polyP提取通过结合到玻璃奶与非常敏感的PolyP激酶/荧光素酶检测系统。 通过使用该程序,我们显着增强了polyP检测的灵敏度,使其可能适用于哺乳动物组织。

【背景】聚磷酸盐(polyP)是一种由多达1,000种无机磷酸盐单体的直链组成的生物聚合物,存在于生命的所有三个领域的细胞中。然而,细菌是唯一已经充分研究了polyP代谢酶的生物。将ATP转化为polyP的细菌polyP激酶(PPK)催化正向和反向反应。虽然polyP的合成显然是细胞中有利的反应,但通过在体外提供足够量的ADP ,该酶可用于从polyP产生ATP,使得基于荧光素酶的ATP检测成为可能(Ault -Riché et al。,1998)。缺乏PPK的细菌在生物膜形成,运动性,持久性和各种应激反应方面存在缺陷,并显示出对次卤酸(即,漂白)应激或磷酸盐饥饿的显着增加的敏感性(图1)(Rao et al。,2009; Gray et ...

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