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Author:
Date:
2013-06-20
[Abstract] Calcium mobilization assay is a cell-based second messenger assay to measure the calcium flux associated with Gq-protein coupled receptor activation or inhibition. The method utilizes a calcium sensitive fluorescent dye that is taken up into the cytoplasm of most cells. In some cell lines in which organic-anion transporters are particularly active (e.g. CHO, HeLa), addition of probenecid, an inhibitor of anion transport, is required for retention of this dye in the cells. The dye binds the calcium released from intracellular store and its fluorescence intensity increases. The change ...
[摘要] 钙动员测定是基于细胞的第二信使测定法，用于测量与Gq蛋白偶联受体激活或抑制相关的钙通量。该方法使用钙敏感荧光染料，其被吸收到大多数细胞的细胞质中。在其中有机 - 阴离子转运蛋白特别活性的一些细胞系（例如CHO，HeLa）中，需要加入丙烯酸，阴离子转运抑制剂，以将该染料保留在细胞中。染料结合从细胞内储存释放的钙，其荧光强度增加。荧光强度的变化与响应感兴趣受体的配体激活释放到细胞质中的细胞内钙的量直接相关。该方案可以应用于表达内源性和瞬时/稳定转染受体的大多数哺乳动物细胞系。该方法足够敏感，可用于低表达系统或感兴趣的靶标的高通量筛选。
注意：该方法不区分Gqα诱导的Ca2 +动员与Gβγ诱导的Ca2 +动员。
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