Cell-Based Potassium Ion Channel Screening Using the FluxOR™ Assay

被引：50

作者：

Beacham, Daniel W. ^{[1
]}

Blackmer, Trillium ^{[1
]}

O'Grady, Michael ^{[1
]}

Hanson, George T. ^{[1
]}

机构：

[1] Life Technol Corp, Invitrogen Mol Probes, Cellular Syst Div, Eugene, OR 97402 USA

来源：

JOURNAL OF BIOMOLECULAR SCREENING | 2010年 / 15卷 / 04期

关键词：

FluxOR (TM); potassium ion channel; high-throughput screening; flux; BacMam; POTENT; EPILEPSY; MUTATION; KCNQ2; SELECTIVITY; ANTAGONIST; INHIBITOR; RECEPTORS;

D O I：

10.1177/1087057109359807

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

FluxOR (TM) technology is a cell-based assay used for high-throughput screening measurements of potassium channel activity. Using thallium influx as a surrogate indicator of potassium ion channel activity, the FluxOR (TM) Potassium Ion Channel Assay is based on the activation of a novel fluorescent dye. This indicator reports channel activity with a large fluorogenic response and is proportional to the number of open potassium channels on the cell, making it extremely useful for studying K+ channel targets. In contrast to BTC-AM ester, FluxOR (TM) dye is roughly 10-fold more thallium sensitive, requiring much lower thallium for a larger signal window. This also means that the assay is carried out in a physiological, normal-chloride saline. In this article, the authors describe how they used BacMam gene delivery to express Kv7.2 and 7.3 (KCNQ), Kir2.1, or Kv11.1 (hERG) potassium ion channels in U2-OS cells. Using these cells, they ran the FluxOR (TM) assay to identify and characterize channel-specific inhibitory compounds discovered within the library (Tocriscreen (TM) Mini 1200 and Sigma Sodium/Potassium Modulators Ligand set). The FluxOR (TM) assay was able to identify several known specific inhibitors of Kv7.2/7.3 or hERG, highlighting its potential to identify novel and more efficacious small-molecule modulators. (Journal of Biomolecular Screening 2010:441-446)

引用

页码：441 / 446

页数：6

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