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Innoprot

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        • DAG-NOMAD GPCR Cell Lines

DAG-NOMAD Muscarinic 5 Cell Line

Reference: P70603


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Data Sheet

A novel M5 Nomad cell line has been developed stably transfecting DAG Nomad biosensor and muscarinic 5 receptor termed M5 into U2OS cell line.

Innoprot DAG-Nomad M5 cell line has been designed to assay compounds or analyze their capability to modulate Muscarinic 5 receptor. When the agonist binds to M5 a G protein is activated, which in turn, triggers a cellular response mediated by second messengers (DAG).


Assay Details


In absence of DAG, the fluorescent biosensor is localized in the cellular membrane. An increase in this second messenger concentration leads to change in the structural folding of DAG-Nomad Biosensor that promotes its cellular relocation in the vesicular trafficking of the cells and a fluorescence intensity increase. Activity is easily quantified on living cells by fluorescence intensity using any microplate reader.

This highly reproducible assay has been validated using oxotremorine as a M5 agonist in both fluorescent intensity and Image analysis platforms. This cell line has been validated measuring DAG increase in the cytosol analyzing Nomad biosensor distribution and fluorescence increase, showing averages higher than of Z´= 0.65 +/- 0.01.


Materials Provided


  • P70203: 2 vials of 3 million stably transfected U2OS Cells expressing M5 and DAG-Nomad Biosensor 
 

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