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HEPES 1M Solution

HEPES溶液

Company: Cytiva
Catalog#: SH30237.01
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Quantitative Measurements of HIV-1 and Dextran Capture by Human Monocyte-derived Dendritic Cells (MDDCs)
Author:
Date:
2016-11-20
[Abstract]  The aim of this protocol is to describe how to measure and quantify the amount of HIV-1 particles and dextran molecules internalized in human monocyte derived dendritic cells (MDDCs), using three different techniques: flow cytometry, quantitative PCR and confocal microscopy. [摘要]  本协议的目的是描述如何使用三种不同的技术:流式细胞术,定量PCR和共聚焦显微镜来测量和量化人类单核细胞衍生树突细胞(MDDCs)中内在的HIV-1颗粒和葡聚糖分子的量。

[背景] 开发此协议是为了评估在人单核细胞衍生的树突细胞中肌动蛋白成核破坏时HIV-1内化的变化。在shRNA筛选后,识别对于HIV-1从树突状细胞转移到T细胞重要的基因,我们观察到肌动蛋白成核的中断导致从富含肌动蛋白的树突到水泡的转变,由于过量的肌动球蛋白收缩。结果,观察到HIV-1转移的减少和由于水泡收缩驱动的巨噬细胞增多引起的HIV-1内化的增加。我们的结论是,肌动蛋白成核和稳定的效应器是维持艾滋病毒1对肌动蛋白丰富的树突和限制其内吞作用,有效转移到T淋巴细胞的关键(Menager和Littman,2016)。

Neutral Red Assay for Murine Norovirus Replication and Detection in a Mouse
Author:
Date:
2013-04-05
[Abstract]  Neutral red (NR) is a dye that must be actively imported into the cell, and, therefore, the dye has been used for decades to selectively stain living cells. In addition, NR can also be incorporated into virus particles, although the mechanism behind this is poorly understood. Once encapsulated into the virion, NR, a light sensitive dye, can be photoactivated to inactivate the virus. The proposed mechanism explaining this observation is that activation of NR allows the dye to cross-link viral genome to viral capsid and thus preventing viral uncoating and infection. To study the early events of murine norovirus (MNV)-host interaction, light-sensitive NR-containing MNV is used to distinguish between input virus (i.e., NR-containing virus) and replicated virus (i.e., ... [摘要]  中性红(NR)是必须主动导入细胞的染料,因此,染料已经使用数十年来选择性地染色活细胞。此外,NR也可以被并入病毒颗粒中,虽然其背后的机制知之甚少。一旦包封入病毒粒子中,NR(一种光敏染料)可以被光活化以使病毒失活。解释这种观察的提出的机制是NR的活化允许染料将病毒基因组交联病毒衣壳,从而防止病毒脱壳和感染。为了研究鼠诺如病毒(MNV) - 宿主相互作用的早期事件,使用光敏感的含NR的MNV来区分输入病毒(即含有NR的病毒)和复制的病毒( >即,无NR的病毒)。该方案描述了将NR掺入MNV衣壳中以及这些病毒颗粒用于通过标准噬菌斑测定在小鼠和组织培养物中检测病毒复制的用途。相同的技术也用于研究脊髓灰质炎病毒复制(1-3)。因此,存在该技术可用于另外的非包膜病毒的潜力。然而,这必须在逐案基础上进行测试,因为关于猫杯状病毒的未公开数据表明不是所有的病毒都能够将NR稳定地掺入其衣壳中(J.Parker,personal communication)。

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