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Corning® 50 mL PP Centrifuge Tubes, Conical Bottom with CentriStarTM Cap, Bulk Packed, Sterile

Corning ® 50mL PP离心管,锥形底部,带CentriStar TM

Company: Corning
Catalog#: 430829
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Liposome Flotation Assay for Studying Interactions Between Rubella Virus Particles and Lipid Membranes
Author:
Date:
2018-08-20
[Abstract]  Rubella virus (RuV) is an enveloped, positive-sense single-stranded RNA virus that is pathogenic to humans. RuV binds to the target cell via the viral envelope protein E1, but the specific receptor molecules on the target cell are yet to be fully elucidated. Here, we describe a protocol for liposome flotation assay to study direct interactions between RuV particles and lipid membranes in a qualitative manner. Interactions are examined by a Nycodenz density gradient fractionation using UV-inactivated RuV particles and fluorescent-labeled liposomes consisting of pure lipids. Fractionated RuV particles are detected using standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blot analysis for viral proteins. On the Nycodenz gradient, RuV particles ... [摘要]  风疹病毒(RuV)是一种包膜的正义单链RNA病毒,对人类具有致病性。 RuV通过病毒包膜蛋白E1与靶细胞结合,但靶细胞上的特异性受体分子尚未完全阐明。在这里,我们描述了脂质体浮选测定的方案,以定性方式研究RuV颗粒和脂质膜之间的直接相互作用。使用UV-灭活的RuV颗粒和由纯脂质组成的荧光标记的脂质体通过Nycodenz密度梯度分级检查相互作用。使用标准十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS-PAGE)检测分级的RuV颗粒,然后对病毒蛋白进行Western印迹分析。在Nycodenz梯度上,与未结合的RuV颗粒相比,与脂质体结合的RuV颗粒转移至较低密度的部分。使用该方案,我们提供了令人信服的证据,即在中性pH下以钙依赖性方式,RuV颗粒与某些细胞类型中含有鞘磷脂(SM)和胆固醇的脂质膜结合。

【背景】 风疹病毒是“风疹”的致病因子,“风疹”是一种急性且相对轻微的全身性感染和“先天性风疹综合征”,一种导致严重出生缺陷的转胎胎儿感染(Hobman,2013)。阐明RuV进入的分子机制对于了解病毒病理学和帮助开发抗RuV药物是必不可少的。虽然以前的研究表明宿主细胞的膜脂质作为RuV受体(Mastromarino et al。,1989和1990; DuBois et ...

Protocol for the Isolation and Super-resolution dSTORM Imaging of RyR2 in Cardiac Myocytes
Author:
Date:
2018-08-05
[Abstract]  Since its inception, super-resolution microscopy has played an increasingly important role in the discovery and characterization of nanoscale biological structure. dSTORM, which is one of the most commonly applied methods, relies on stochastic photoswitching of fluorophores to recreate a super-resolution image. The cardiac field has particularly benefitted from the application of this technique, as it has enabled sub-diffraction-limit visualization of calcium release units (CRUs) and the fundamental structures that trigger contraction. Acquisition of such images requires careful, reproducible sample preparation, and consistent imaging conditions maintained for the duration of the experiment. Here we present standardized methods for the production of dSTORM images of the Ca2+ ... [摘要]  自成立以来,超分辨率显微镜在纳米级生物结构的发现和表征中发挥着越来越重要的作用。 dSTORM是最常用的方法之一,它依赖于荧光团的随机光切换来重建超分辨率图像。心脏场特别受益于该技术的应用,因为它已经实现了钙释放单元(CRU)的子衍射极限可视化和触发收缩的基本结构。获取这些图像需要仔细,可重复的样品制备,并且在实验期间保持一致的成像条件。在这里,我们提出了生产心肌细胞中Ca 2 + 释放通道Ryanodine Receptor type-2(RyR2)的dSTORM图像的标准化方法。所提出的方案特别关注涉及原发性心肌细胞分离,样品制备和成像的步骤,其中提供了针对实验溶液和显微镜设置的细节。本讨论之后是各种分析技术的概述,以识别集群和CRU中的RyR2组织

【背景】近年来,超分辨率显微镜的普及率迅速提高。已经描述了各种超分辨率技术,其使光学分辨率远低于光的衍射极限,在某些情况下接近可通过电子显微镜获得的光学分辨率。总之,这些技术的出现导致了纳米级生物结构,结构域和蛋白质相互作用的新研究的爆炸式增长。一种流行的超分辨率技术是直接随机光学显微镜(dSTORM),与标准共聚焦显微镜相比,它将相对简单的样品处理的优势与分辨率提高了约10倍(van de Linde ...

Brain Tissue Culture of Per2::Luciferase Transgenic Mice for ex vivo Bioluminescence
Author:
Date:
2018-07-05
[Abstract]  In circadian research, it is essential to be able to track a biological rhythm for several days with the minimum perturbation for the organisms or tissues. The use of transgenic mice lines, in which the luciferase reporter is coupled to a molecular clock protein (here PERIOD2), gives us the opportunity to follow the circadian activity in different tissues or even single clock cells for days without manipulation. This method creates sections using a mouse brain matrix, which allows us to obtain several brain samples quickly at a single time point. [摘要]  在昼夜节律研究中,能够以最小的生物或组织扰动跟踪生物节律数天是至关重要的。 使用转基因小鼠系,其中荧光素酶报告基因与分子钟蛋白(此处为PERIOD2)偶联,使我们有机会在不经操作的情况下跟踪不同组织或甚至单个时钟细胞中的昼夜节律活动数天。 该方法使用鼠标脑矩阵创建切片,这允许我们在单个时间点快速获得几个脑样本。

【背景】昼夜节律是大约24小时循环的行为或分子变化,并且在没有任何外部线索的情况下持续。在哺乳动物中,运动活动,体温和激素释放是昼夜节律的实例,其在位于下丘脑的视交叉上核(SCN)时钟的控制下。 SCN细胞保持内源性节律的能力是由于时钟基因表达的正负循环组成的分子机制:首先,CLOCK和BMAL1蛋白异二聚化通过E-box位点激活不同基因的转录关于基因如 period ( Per1-3 )和隐花色素( Cry1-2 ; Takahashi)的启动子 et al。,2008)。然后,PERIOD和CRYPTOCHROME的蛋白质异二聚化并返回到细胞核以防止BMAL1与E-Box结合。因此,PERIOD和CRYPTOCHROME抑制其自身的转录(Takahashi et al。,2008)。第二个环由类视黄醇相关的孤儿受体(ROR)和Rev-Erb组成:ROR蛋白激活 Bmal1 基因,而REV-ERB蛋白抑制它通过 ROR反应 Bmal1 ...

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