Cell-based Calcium Assay for Medium to High Throughput Screening of TRP Channel Functions using FlexStation 3

Jialie Luo; Yingmin Zhu; Michael X. Zhu; Hongzhen Hu

doi:10.3791/3149

Cell-based Calcium Assay for Medium to High Throughput Screening of TRP Channel Functions using F...

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Cell-based Calcium Assay for Medium to High Throughput Screening of TRP Channel Functions using FlexStation 3

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07:26 min

August 17, 2011

DOI: 10.3791/3149-v

Jialie Luo¹, Yingmin Zhu¹, Michael X. Zhu¹, Hongzhen Hu¹

¹Department of Integrative Biology and Pharmacology,The University of Texas Health Science Center at Houston

This video provides a detailed protocol for studying the pharmacological profile of human TRPA1 channels using FlexStation 3. The protocol covers details of cell preparation, dye loading and operation of the microplate reader, FlexStation 3.

The overall goal of this procedure is to demonstrate the application of Flex Station three to study the pharmacological profile of the transient receptor potential or trip a one channel. This is accomplished by first preparing cells that express trip a one channels in 96. Well microtiter plates.

After washing the cells in a microplate washer, the cells are loaded with the calcium sensitive fluorescent dye. Flu oh 4:00 AM a cell loading period is then followed by several additional washes. The next step of the procedure is to prepare the compound plate, including a trip, a one agonist and a putative trip, a one antagonist.

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