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Fixation buffer from BD Fix/Perm kit

固定/渗透溶液试剂盒(RUO)

Company: BD
Catalog#: 554714
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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)。

In vivo BrdU Incorporation Assay for Murine Hematopioetic Stem Cells
Author:
Date:
2013-11-05
[Abstract]  Bromodeoxyuridine (BrdU) is a thymidine analog that is incorporated into DNA during the S-phase of the cell cycle. As such, BrdU incorporation can be used to quantify the number of cells that are in S-phase in the time period during which BrdU is available. The following protocol describes an in vivo BrdU incorporation assay as a measure of cell proliferation in adult murine hematopioetic stem cells (HSCs). Specifically, BrdU incorporation was analyzed for long-term HSCs (LT-HSCs, Lin-Sca-1+c-Kit+CD34-CD135-), Short-term HSCs (ST-HSCs, Lin-Sca-1+c-Kit+CD34+CD135-) and multipotent progenitors (MPPs, Lin-Sca-1+c-Kit+CD34+CD135+) population.
[摘要]  溴脱氧尿苷(BrdU)是在细胞周期的S期期间掺入DNA的胸苷类似物。 因此,BrdU掺入可用于定量在BrdU可用的时间段内处于S期的细胞的数量。 以下方案描述了体内BrdU掺入测定作为成体鼠造血干细胞(HSC)中细胞增殖的量度。 具体地,分析BrdU掺入的长期HSC(LT-HSC,Lin-Sca-1 + c-Kit + CD34 - CD135 - ),短期HSC(ST-HSC,Lin-Sca-1 + c-Kit + CD34 和多能祖细胞(MPP,Lin-Sca-1 + c-Kit + CD34 + > CD135 + )群体

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