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Sodium dodecyl sulfate

十二烷基硫酸钠

Company: Sigma-Aldrich
Catalog#: L3771
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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 ...

Identifying Protein Interactions with Histone Peptides Using Bio-layer Interferometry
Author:
Date:
2018-09-20
[Abstract]  Histone post-translational modifications (PTMs) regulate numerous cellular processes, including gene transcription, cell division, and DNA damage repair. Most histone PTMs affect the recruitment or exclusion of reader proteins from chromatin. Here, we present a protocol to measure affinity and interaction kinetics between histone peptides and the recombinant protein using Bio-layer interferometry. [摘要]  组蛋白翻译后修饰(PTM)调节许多细胞过程,包括基因转录,细胞分裂和DNA损伤修复。 大多数组蛋白PTM影响从染色质中募集或排除读取蛋白。 在这里,我们提出了一个协议,使用生物层干涉测量法测量组蛋白肽和重组蛋白之间的亲和力和相互作用动力学。

【背景】真核染色质结构大致分为常染色质和异染色质(Cheung和Lau,2005),异染色质结构根据组蛋白翻译后修饰(PTM)的组合进一步细分。这些PTM不仅改变染色质构象,还在基因表达和蛋白质募集中建立直接调节作用(Felsenfeld和Groudine,2003; Allshire和Madhani,2017)。组蛋白PTM的无数组合 - 包括乙酰化,磷酸化,甲基化,泛素化,生物素化,SUMO化和脯氨酸异构化,统称为“组蛋白标记” - 可以被发现,特别是在从核小体核心突出的非结构化N末端尾部( Guetg和Santoro,2012)。这些PTM通过不同“读者”或效应蛋白的活动调节许多细胞过程,包括基因转录,细胞分裂和DNA损伤修复(Suganuma和Workman,2011)(Musselman et al。, 2012)。因此,已经做出很大努力来识别读者的组蛋白修饰。

使用常规方法(例如,表面等离子体共振[SPR]和SPR成像[SPRi]生物传感器)研究读取蛋白与其靶蛋白PTM之间的相互作用通常需要大量底物或复杂的多步实验方法并且由于各种方法特定的限制而变得复杂。这些问题排除了量化相互作用强度的简便性和准确性(Phizicky和Fields,1995; ...

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