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NTSR1 Nomad Mpx Cell Line

Reference: P70746


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Data Sheet

A novel NTSR1 Nomad Mpx cell line has been developed stably transfecting red fluorescent b-arrestin Nomad biosensor, green fluorescent Ca++ Nomad biosensor and label free human neurotensin receptor 1 termed NTSR1  into U2OS cell line.

Innoprot NTSR1 Nomad Mpx cell line has been designed to assay compounds or analyze their capability to modulate neurotensin receptor 1. When the agonist binds to NTSR1 a G protein is activated, which in turn, triggers a cellular response mediated by Ca++ 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 in a single assay.


Assay Details


In absence of Ca++, the green fluorescent biosensors are localized in the cellular membrane. An increase in the Ca++ concentration leads to change in the structural folding of the green 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 red fuorescent biosensor interacts with the recruited b-arrestin, leading to change in the structural folding of the red Nomad Biosensor promotes its cellular relocation 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 Neurotensin as a NTSR1 agonist in a High Content Analysis (HCA) Platform for image analysis and in any microplate reader for fluorescence intensity. This cell line has been validated measuring b-arrestin recruitment and Ca++  increase in the cytosol analyzing both Nomad biosensor intensity and distribution within the cell, showing an averages higher than Z´= 0.75 +/- 0.01 in all determinations.


Materials Provided


  • P70746: 2 vials of 3 million stably transfected U2OS Cells expressing NTSR1, Ca++ Nomad Biosensor & b-arrestin Nomad biosensor
   

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