{{'Search' | translate}}
 

Open-Top Thinwall Ultra-Clear Tube (13.2 ml), 14 x 89 mm

管,薄壁,超透明TM ,13.2mL,14×89mm

Company: Beckman Coulter
Catalog#: 344059
Bio-protocol()
Company-protocol()
Other protocol()

Buoyant Density Fractionation of Small Extracellular Vesicle Sub-populations Derived from Mammalian Cells
Author:
Date:
2020-08-05
[Abstract]  Small extracellular vesicles (sEVs) encompass a variety of distinct vesicles that are secreted to the extracellular space. Many methodologies currently used for EV isolation (e.g., differential ultracentrifugation concluding in a high-speed pellet, precipitation by macromolecular crowding agents or size excusion chromatography–SEC) do not fractionate distinct sEV sub-populations. Samples obtained by the aforementioned methods are usually used for characterization and physiological studies. However the fraction that contains the molecule of interest or is the carrier of a specific activity is unknown. Therefore isolating distinct sEV sub-populations is critical to understand EV function. The goal of this procedure is to purify distinct sEV sub-populations based on slight ... [摘要]  [摘要] 小细胞外小泡(sEVs)包括分泌到细胞外空间的各种不同的小泡。目前用于EV分离的许多方法(例如,高速颗粒中的差速超速离心、大分子拥挤剂沉淀或尺寸排除色谱法)没有分离不同的sEV亚群。通过上述方法获得的样品通常用于表征和生理学研究。然而,包含感兴趣分子或特定活性载体的部分是未知的。因此,分离不同的sEV亚群对于理解EV功能至关重要。该程序的目的是基于它们浮力密度的微小差异来纯化不同的sEV亚群。此外,该技术还允许从高速颗粒中共同分离的无囊泡RNA蛋白复合物中或通过使用拥挤剂来纯化sEVs。该方案描述了用于收集sEV的哺乳动物细胞的培养、sEV沉淀、sEV亚群的浮力密度分馏和sEV标记的免疫印迹。该方法可用于分离由多种哺乳动物细胞产生的不同的sEV亚群。

[] ...

Isolation of Lipid Rafts from Cultured Mammalian Cells and Their Lipidomics Analysis
Author:
Date:
2020-07-05
[Abstract]  Lipid rafts are distinct liquid-ordered domains of plasma membranes of most eukaryotic cells providing platform for signaling pathways. Lipid composition of rafts is critical for their structural integrity and for regulation of signaling pathways originating from rafts. Here we provide a protocol to isolate lipid rafts from cultured human and animal cells and comprehensively analyse their lipid composition. [摘要]  [摘要]脂质筏是大多数真核细胞浆膜中独特的液态有序域,为信号传导途径提供平台。脂质筏的脂质组成对其结构的完整性和对源自脂质筏的信号传导途径的调节至关重要。在这里,我们提供了一个协议,从培养的人和动物细胞分离脂质筏,并全面分析其脂质组成。
关键词脂质筏,脂质,脂质组学,胆固醇,膜

[背景]脂质筏是细胞膜中独特的富含胆固醇和鞘氨醇的结构,浸入液态无序的周围膜中(Lingwood和Simons,2010年)。它们提供了一个"坚实"的平台,在空间上组织各种信号传导途径的元素,以及外泌和内吞机制。脂筏的功能特性由脂筏脂质和蛋白质之间的相互作用决定。脂筏脂质组成的变化,无论是生理性的还是病理性的,都会对源自脂筏的途径的活性产生惊人的后果,并代表了其调节的一个重要的、未被充分重视的层面。脂质筏的分离因其动态性和异质性而变得复杂。基于它们对洗涤剂的抵抗力的筏子分离的方法一直受到批评(高斯 et al., 2005)。在这里,我们描述了一个无洗涤剂的方法,用于分离脂筏和建立他们的脂质组成,使用综合脂质组学分析。最近发表了一个使用该协议的例子(Mukhamedova等人,2019)。

第一部分:脂质筏的分离

下面描述的协议是用来从RAW ...

Isolation and Characterization of Exosomes from Mouse Feces
Author:
Date:
2020-04-20
[Abstract]  Exosomes secreted by colonic epithelial cells are present in feces and contain valuable epigenetic information, such as miRNAs, proteins, and metabolites. An in-depth study of this information is conducive to the diagnosis or treatment of relevant diseases. A crucial prerequisite of such a study is to establish an efficient isolation method, through which we can obtain a relatively more significant amount of exosomes from feces. This protocol is designed to effectively isolate a large number of exosomes from contaminants and other particles in feces by a combined method with fast filtration and sucrose density gradient ultracentrifugation. Exosomes generated by this method are suitable for further RNA, protein, and lipid analysis. [摘要]  [摘要] 外来体分泌的结肠上皮细胞是否存在,在粪便和包含有价值的表观遗传信息,例如miRNA,蛋白质和代谢。一个在深入研究这个信息,有利于诊断和治疗相关疾病的一种重要前提这项研究的目的是建立一种有效的分离方法,通过该方法我们可以从粪便中获得相对大量的外泌体。该方案旨在通过组合方法有效地从粪便中的污染物和其他颗粒中分离出大量外泌体通过快速过滤和蔗糖密度梯度超速离心。这种方法产生的外泌体适用于进一步的RNA,蛋白质和脂质分析。

[背景] 结肠外泌体由结肠上皮细胞分泌到管腔中,并沿大肠转运并存在于粪便中。这些外泌体的脂质双层结构可防止复杂条件下封装的生物分子(如miRNA)的降解(由于粪便)(古贺等人,2011 ; 邓等人,2013 )。该保护功能外体是非常有用的,因为这些受保护的内容可以用来诊断疾病,如溃疡性结肠炎和结肠癌症。重要的是,重新设计的外泌体也可以有效地将治疗性生物分子递送至某些特定的疾病靶标,而不会对宿主产生免疫毒性(Sun 等人,2010; Johnsen 等人,2014; Wang 等人,2016; Kim和Kim,2018 )。

迄今为止,已成功地从血液(Wu 等人,2017 ),尿液(Knepper和Pisitkun,2007; Motamedinia 等人,2016 ),培养细胞(Yeo 等人,2013 ...

Comments