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50 ml conical tubes

Corning ® 50mL PP离心管,锥形底部带插塞密封帽

Company: Corning
Catalog#: 430290
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Tracking Endocytosis and Intracellular Trafficking of Epitope-tagged Syntaxin 3 by Antibody Feeding in Live, Polarized MDCK Cells
[Abstract]  The uptake and trafficking of cell surface receptors can be monitored by a technique called ‘antibody-feeding’ which uses an externally applied antibody to label the receptor on the surface of cultured, live cells. Here, we adapt the traditional antibody-feeding experiment to polarized epithelial cells (Madin-Darby Canine Kidney) grown on permeable Transwell supports. By adding two tandem extracellular Myc epitope tags to the C-terminus of the SNARE protein syntaxin 3 (Stx3), we provided a site where an antibody could bind, allowing us to perform antibody-feeding experiments on cells with distinct apical and basolateral membranes. With this procedure, we observed the endocytosis and intracellular trafficking of Stx3. Specifically, we assessed the internalization rate of Stx3 from the ... [摘要]  细胞表面受体的摄取和贩卖可以通过被称为“抗体 - 进纸”技术,其在外部使用上应用的抗体来标记培养的活细胞的表面上的受体进行监测。在这里,我们适应传统的抗体喂养实验极化上皮细胞(Madin-Darby犬肾)生长在透性transwell支持。通过添加两个串联胞外Myc表位标签的SNARE蛋白突触3(Stx3)的C末端,我们提供了一种站点,其中与抗体可以结合,使我们能够用不同的顶端和基底外侧膜对细胞进行抗体 - 喂养实验。通过这个程序,我们观察到Stx3的内吞和细胞内运输。具体而言,我们评估从基底外侧膜Stx3的内在化速率和在时间和空间上使用固定在不同时间点的细胞免疫荧光显微镜随后的内吞观察路径。为此,介绍了以下可追踪单层的贩运行为:来自限制膜的靶蛋白。

【背景】SNARE蛋白突触3(Stx3)是已知的建立在极化上皮细胞顶端 - 基底极性(低等人,1996年Delgrossi 等人,1997年Weimbs 等人,1997;低等人,1998;黎等人,2002;低等人

一个典型的12孔细胞培养皿的阱内的transwell小室的聚碳酸酯膜的细胞培养插入物的图1示意图。斯特朗细胞是在膜的顶部和培养将形成紧密的单层做密封关聚碳酸酯膜将顶端介质隔室与基底外侧介质隔室分开。 ...

Organotypic Brain Cultures: A Framework for Studying CNS Infection by Neurotropic Viruses and Screening Antiviral Drugs
[Abstract]  According to the World Health Organization (WHO), at least 50% of emerging viruses endowed with pathogenicity in humans can infect the Central Nervous System (CNS) with induction of encephalitis and other neurologic diseases (Taylor et al., 2001; Olival and Daszak, 2005). While neurological diseases are progressively documented, the underlying cellular and molecular mechanisms involved in virus infection and dissemination within the CNS are still poorly understood (Swanson and McGavern, 2015; Ludlow et al., 2016). For example, measles virus (MeV) can infect neural cells, and cause a persistent brain infections leading to lethal encephalitis from several months to years after primary infection with no available treatment (Reuter and Schneider-Schaulies, 2010; Laksono et ... [摘要]  根据世界卫生组织(WHO)的统计,至少有50%的新发病毒具有致病性,可感染中枢神经系统(CNS),并诱发脑炎和其他神经系统疾病(Taylor et al。 >,2001; Olival和Daszak,2005)。虽然神经系统疾病逐渐被记录下来,但涉及病毒感染和在CNS内传播的潜在细胞和分子机制仍然知之甚少(Swanson and McGavern,2015; Ludlow等人,2016)。例如,麻疹病毒(MeV)可以感染神经细胞,并在原发感染后几个月至数年导致持续的脑感染,导致致死性脑炎,而没有可用的治疗(Reuter和Schneider-Schaulies,2010; Laksono等人。,2016)。器官型脑文化(OBC)是病毒学领域的一个合适的模型,以更好地理解中枢神经系统感染。实际上,它不仅可以研究中枢神经系统内嗜神经病毒的感染和传播,而且还可以作为创新性抗病毒策略或分子的筛选模型,如我们最近发表的有关融合抑制肽和HSP90伴侣蛋白活性抑制剂的研究, 17-DMAG(Welsch等人,2013; ...

Isolation and Purification of Schwann Cells from Spinal Nerves of Neonatal Rat
[Abstract]  Primary cultured Schwann cells (SCs) are widely used in the investigation of the biology of SC and are important seed cells for neural tissue engineering. Here, we describe a novel protocol for harvesting primary cultured SCs from neonatal Sprague-Dawley (SD) rats. In the present protocol, dissociated SCs are isolated from the spinal nerves of neonatal rats and purified by the treatment of cytosine arabinoside (AraC). [摘要]  原代培养的雪旺氏细胞(SCs)广泛用于SC的生物学研究,是神经组织工程的重要种子细胞。 在这里,我们描述了从新生Sprague-Dawley(SD)大鼠中收获原代培养的SC的新方案。 在本方案中,解离的SCs从新生大鼠的脊神经中分离并通过治疗阿糖胞苷(AraC)进行纯化。
【背景】SC是周围神经系统(PNS)的神经胶质细胞。主要SCs的分离和纯化是研究SC生物学的关键步骤。此外,纯化的原代培养的SC是用于神经组织工程的重要种子细胞。迄今为止,已经基于Brockes(Brockes等人,1979)的方法报道了培养SC的各种方法。通过报道的方法,坐骨神经主要用于SC隔离,因为它们尺寸大并且可以容易地获得。然而,坐骨神经的SC容易被成纤维细胞污染,因为结缔组织难以清除。特别是神经管和神经束是成纤维细胞的主要来源。没有特殊处理,污染的成纤维细胞比SCs快得多,并且将很快成为培养物中主要的细胞。在过去几十年中,已经开发出许多用于从污染成纤维细胞中分离SC的纯化方法。这些纯化方法的细节包括单一或组合的抗有丝分裂处理(Wood,1976),抗体介导的细胞溶解(Brockes等人,1979),免疫选择(Assouline等人,,1983; ...