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Innoprot

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        • Multiplex NOMAD GPCR Cell Lines

CB1 Nomad Cell Line

Reference: P70727


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Data Sheet

A novel CB1 Nomad Mpx cell line has been developed stably transfecting green fluorescent b-arrestin Nomad biosensor, red fluorescent cAMP Nomad biosensor and label free human cannabinoid receptor 1 termed CB1 or CNR1 into HEK293 cell line.

Innoprot CB1 Nomad Mpx cell line has been designed to assay compounds or analyze their capability to modulate Cannabinoid Receptor 1. When the agonist binds to CB1 a G protein is activated, which in turn, triggers a cellular response mediated by cAMP and at the same time β-arrestin-dependent mechanisms can be also activated. Using Mpx Nomad cell lines, it is possible to monitor both activation pathways at the same time in a single assay.


Assay Details


In absence of cAMP, the red fluorescent biosensors are localized in the cellular membrane. An increase in the Ca++ concentration leads to change in the structural folding of the red Nomad Biosensor that promotes its cellular relocation in the vesicular trafficking of the cells. Activity is easily quantified on living cells by both, image analysis of cytoplasmatic granularity changes and fluorescence intensity changes.

Furthemore, the green fuorescent b-arrestin biosensor interacts with the activated GPCR, leading to change in the structural folding of the red Nomad Biosensor. The binding of b-arrestin with the activated GPCR promotes the cellular relocation of the biosensor in the vesicular trafficking of the cells. B-arrestin recruitment is easily quantified on living cells by both, image analysis of cytoplasmatic granularity changes and fluorescence intensity changes.

This highly reproducible assay has been validated using CP55940 as a CB1 agonist in a microplate reader for fluorescence intensity. This cell line has been validated measuring b-arrestin recruitment and cAMP increase in the cytosol analyzing both Nomad biosensor intensity and distribution within the cell, showing an averages higher than Z´= 0.70 +/- 0.01 in all determinations.


Materials Provided


  • P70727: 2 vials of 3 million stably transfected HEK293 Cells expressing CB1, cAMP Nomad Biosensor & b-arrestin Nomad biosensor
   

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