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Dulbecco’s Modified Eagle Medium/Ham’s F-12

DMEM / F-12

Company: Thermo Fisher Scientific
Catalog#: 31331093
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Carboxyfluorescein Dye Uptake to Measure Connexin-mediated Hemichannel Activity in Cultured Cells
Author:
Date:
2021-02-05
[Abstract]  

Connexins are membrane bound proteins that facilitate direct and local paracrine mediated cell-to-cell communication through their ability to oligomerise into hexameric hemichannels. When neighbouring channels align, they form gap-junctions that provide a direct route for information transfer between cells. In contrast to intact gap junctions, which typically open under physiological conditions, undocked hemichannels have a low open probability and mainly open in response to injury. Hemichannels permit the release of small molecules and ions (approximately 1kDa) into the local intercellular environment, and excessive expression/activity has been linked to a number of disease conditions. Carboxyfluorescein dye uptake measures functional expression of hemichannels, where increased ... [摘要]  [摘要]连接蛋白是有助于直接和局部旁分泌通过自己的能力介导的细胞-细胞通信膜结合蛋白寡聚成六聚半通道。当相邻通道对齐时,它们形成间隙连接,为单元之间的信息传输提供直接路径。与通常在生理条件下打开的完整的间隙连接相反,未对接的半通道的打开概率较低,主要是在受伤时打开。半通道允许小分子和离子(约1kDa)释放到局部细胞间环境中,并且过度表达/活性 与许多疾病状况有关。羧基荧光素染料的摄取量度了半通道的功能性表达,其中增加的半通道活性/功能反映了增加的负荷。该技术依赖于通过开放性半通道对不可渗透膜的荧光示踪剂的吸收,可用于比较在不同条件下(例如对照与疾病)培养的细胞单层之间的通道活性。其他技术(例如生物素化和电生理学)可以分别测量细胞表面表达和半通道开放可能性,但是,羧基荧光素的摄取提供了一种简单,快速且经济高效的方法来测定多种细胞类型的体外半通道活性。


图形摘要:

使用染料吸收量度半通道活性

[背景]连接蛋白(CX )是一体的跨膜蛋白,其寡聚到连接子在细胞表面。连接子与相似的六聚体蛋白复合物停靠在相邻细胞上,形成用于间隙连接细胞间通讯的双向导管(GJIC;Bosco等人,2011 ...

Protocol for T-cell Adhesion Strength on Tumor Cells under Flow Conditions
Author:
Date:
2013-10-20
[Abstract]  This method allows evaluating the relative adhesion strength between T lymphocytes and specific adherent target cells using a shear force in flow chambers. It is based on the measure of the resistance of conjugates formed between T cells and adherent tumor cells to shear stress in a microfluidic system. For this purpose, T cells, stained with a CellTracker probe, are added into flow channels containing a monolayer of adherent target cells and their progressive detachment under a constant shear stress is then recorded using a fluorescent microscope. [摘要]  该方法允许使用流动室中的剪切力评估T淋巴细胞和特异性粘附靶细胞之间的相对粘附强度。 其基于在微流体系统中T细胞和粘附的肿瘤细胞之间形成的缀合物对剪切应力的抗性的测量。 为此,将用CellTracker探针染色的T细胞加入含有单层粘附靶细胞的流动通道中,然后使用荧光显微镜记录在恒定剪切应力下的其逐渐分离。

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