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Tube, Thinwall, Ultra-ClearTM, 13.2 mL, 14 x 89 mm (qty. 50)

管,薄壁,超透明TM ,13.2mL,14×89mm

公司名称: Beckman Coulter
产品编号: 344059
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Isolation, Purification and Characterization of Exosomes from Fibroblast Cultures of Skeletal Muscle
Author:
Date:
2020-04-05
[Abstract]  Exosomes are dynamic nanovesicles secreted by virtually all cells and are present in all biological fluids. Given their highly heterogeneous content exosomes have been implicated in many physiological and pathological processes that they exert by influencing cell-cell and cell-ECM communication. In recent years an increasing number of methods have been established for the purification and characterization of exosomes. These include ultracentrifugation, ultrafiltration, size exclusion chromatography, immune capture and precipitation using a proprietary polymer. Here, we provide a protocol ... [摘要]  [摘要 ] 外来体是几乎所有细胞分泌的动态纳米囊泡,并存在于所有生物体液中。鉴于它们的异质含量很高,外泌体已牵涉到它们通过影响细胞-细胞和细胞-ECM通讯而发挥的许多生理和病理过程。近年来,已经建立了越来越多的方法外泌体的纯化和表征。其中包括超速离心,超滤,尺寸排阻色谱,免疫捕获和使用专有聚合物的沉淀。在这里,我们提供了基于差分超速离心和蔗糖密度梯度的协议,该协议专门用于从成年小鼠骨骼肌衍生的原代成纤维细胞培养物中分离粗制和超纯外泌体。可以对该协议进行修改和修改,以从各种组织和体液中分离和表征外泌体。


背景 ] ë xosomes是单膜,异质纳米囊泡直径范围从30至150nm,secre 由所有细胞和存在于几乎所有的体液泰德。外泌体中存在的可溶性和膜大分子,mRNA,microRNA的光谱取决于代谢状态以及分泌这些纳米囊泡的细胞的发育起源。由于它们的货物组成,外泌体可以启动接收细胞中的信号传导途径,并参与了发育,免疫和正常组织生理的维持。在神经退行性疾病,纤维化和癌症等疾病条件下,它们被证明可以触发和传播病理刺激(Rackov 等,2018; Gurunathan 等,2019; van de Vlekkert 等,2019)。在这里,我们描述了从成年小鼠腓肠肌(GA)肌肉建立的成纤维细胞培养物中纯化外泌体的方案(van de Vlekkert ...

Isolation, BODIPY Labeling and Uptake of Exosomes in Hepatic Stellate Cells
Author:
Date:
2017-12-05
[Abstract]  Exosomes have emerged as an important mediator of intercellular communication. They are present in extracellular milieu and therefore, easily accessible by neighboring or distant cells. They carry mRNA, microRNAs and proteins within their vesicles and once internalized by recipient cells; they can modulate multiple signaling pathways with pleiotropic effects from inducing antiviral state to disease progression. We have previously shown that hepatitis C virus (HCV) infected hepatocytes or hepatoma cells harboring genome-length replicon secrete exosomes in culture supernatants. These exosomes ... [摘要]  外来体已经成为细胞间通讯的重要介质。 它们存在于细胞外环境中,因此可以被邻近或远处的细胞轻易获得。 它们在囊泡内携带mRNA,microRNA和蛋白质,并一度被受体细胞内化; 它们可以通过诱导抗病毒状态向疾病进展调节多种信号传导途径,具有多效性。 我们以前已经显示,携带基因组长度复制子的丙型肝炎病毒(HCV)感染的肝细胞或肝癌细胞在培养上清液中分泌外泌体。 这些外来体被肝星状细胞(HSC)摄取,并在HCV感染期间激活它们以诱导纤维化。 在这里,我们描述了外来体分离和肝星状细胞摄取BODIPY标记的外来体的详细的协议。

【背景】外来体是直径为40-150nm的小的膜结合的细胞外囊泡。已在所有体液中鉴定出它们,包括尿,羊水,血清,唾液,母乳,脑脊液,鼻分泌物和组织培养的细胞系的上清液中。外泌体通过转移微小RNA(mRNA),mRNA和蛋白质来介导细胞与细胞之间的通讯。 其可被邻近或远处的细胞摄取,并随后促进受体细胞中的信号传导(Schorey等人,2015)。外泌体富含蛋白质,包括四跨膜蛋白家族成员(CD9,CD81,CD63),热休克蛋白(Hsp60,Hsp70,Hsp90)和多泡体蛋白(annexins,RabGTPases和内体分选复合物,蛋白质)。最近的研究表明,外来体可以影响免疫应答(Li等人,2013; ...

Cell-free Fluorescent Intra-Golgi Retrograde Vesicle Trafficking Assay
Author:
Date:
2017-11-20
[Abstract]  Intra-Golgi retrograde vesicle transport is used to traffic and sort resident Golgi enzymes to their appropriate cisternal locations. An assay was established to investigate the molecular details of vesicle targeting in a cell-free system. Stable cell lines were generated in which the trans-Golgi enzyme galactosyltransferase (GalT) was tagged with either CFP or YFP. Given that GalT is recycled to the cisterna where it is located at steady state, GalT-containing vesicles target GalT-containing cisternal membranes. Golgi membranes were therefore isolated from GalT-CFP expressing cells, ... [摘要]  高尔基体内的逆行囊泡运输被用来运送和分类高尔基酶到适当的池内位置。建立了一个检测方法来研究无细胞系统中囊泡靶向的分子细节。生成了稳定的细胞系,其中反式 - 高尔基酶半乳糖基转移酶(GalT)用CFP或YFP标记。考虑到GalT被循环到稳定状态的小池中,含有GalT的囊泡将靶向含有GalT的池内膜。因此从表达GalT-CFP的细胞分离高尔基体膜,而从GalT-YFP表达细胞制备囊泡。在胞质溶胶和能量再生混合物的存在下,在37℃孵育CFP标记的高尔基体和YFP标记的囊泡,与在冰上孵育相比,在混合物的荧光成像后CFP-YFP共定位显着增加。该测定被验证需要能量,蛋白质和生理学重要的运输组分,如Rab GTP酶和保守寡聚体高尔基体系复合物。该测定法可用于调查影响高尔基体运输机器的生理和病理变化,特别是囊泡束缚。

【背景】细胞内囊泡靶向的分子机制对于解释糖基化稳态,神经递质释放,信号受体的调节和营养摄取等方面的解释是重要的(Ungar和Hughson,2003; ...

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