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Fluo-4

Fluo-4

Company: Thermo Fisher Scientific
Catalog#: F14201
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Fluorescent Measurement of Synaptic Activity Using SynaptopHluorin in Isolated Hippocampal Neurons
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Date:
2014-12-05
[Abstract]  This protocol comprises the entire process of fluorescent measurement of vesicle recycling using the probe SynaptopHluorin, a pH-dependent GFP variant whose fluorescence increases at the synapse upon vesicle release due to fluorescence quenching in acidic vesicles. This technique provides a genetic tool to monitor synaptic vesicle recycling in real time in cultured hippocampal neurons. [摘要]  该协议包括使用探针SynaptopHluorin的荧光测量的荧光测量的整个过程,所述探针是pH依赖性GFP变体,其荧光在囊泡释放时由于酸性囊泡中的荧光猝灭而在突触上增加。 这种技术提供了一种遗传工具来监测突触小泡实时回收培养的海马神经元。

Calcium Mobilisation Assay in Response to Chemokine Stimulation
Author:
Date:
2012-05-05
[Abstract]  This assay is used to measure calcium mobilization in lymphocytes (either in primary cells or cell lines) in response to chemokine stimulation using ratiometric analysis. It has also been used for measuring TCR mediated calcium flux. In addition, the same labeling procedure with the addition of brilliant black (a quenching agent) (Sigma-Aldrich, catalog number: 211842) in the loading buffer (at 100 μM) allows for quantification using the FLIPR system on poly-D-lysine plates. Probenicid is an anion-exchange protein inhibitor and prevents the extrusion of the dyes by organic ion transporters. [摘要]  该测定用于使用比率分析测量响应趋化因子刺激的淋巴细胞(原代细胞或细胞系)中的钙动员。 它也被用于测量TCR介导的钙通量。 此外,在加样缓冲液(100μM)中加入亮黑(淬灭剂)(Sigma-Aldrich,目录号:211842)的相同标记方法允许使用FLIPR系统定量聚D-赖氨酸 板。 丙磺舒是阴离子交换蛋白抑制剂,并通过有机离子转运蛋白防止染料的挤出。

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