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Innoprot

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        • cAMP-NOMAD GPCR Cell Lines

M4 Nomad Cell Line

Reference: P70545


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Data Sheet

A novel M4 Nomad cell line has been developed stably transfecting PKA Nomad biosensor and Muscarinic 4 receptor termed M4R into U2OS cell line.

Innoprot´s red PKA-Nomad‐CHRM4 cell line has been  designed  to  assay  compounds  or  analyze  their  capability  to  modulate Muscarinic Acetylcholine Receptor M4. When an agonist binds to CHRM4, Gi/o is activated, which  in  turn,  triggers  a  cellular  response  involving  PKA  activation.  Activated  PKA interacts with PKA-Nomad Biosensor, increasing its fluorescence intensity.


Assay Details


In basal conditions, the fluorescent biosensor is localized in the cellular membrane. After receptor activation, PKA is activated and this activated PKA leads to change in the structural folding of Nomad Biosensor that promotes its cellular relocation in the vesicular trafficking of the cells. Activity is easily quantified on living cells by fluorescence intensity but also by image analysis of cytoplasmatic granularity changes.

This highly reproducible assay has been validated using oxitremorine as a M4R agonist in a microplate reader. This cell line has been validated measuring PKA activation analyzing both fluorescence intensity and Nomad biosensor distribution within the cell, showing an average of Z´= 0.81 +/- 0.01.


Materials Provided


  • P70545: 2 vials of 3 million stably transfected U2OS Cells expressing M4R and Nomad Biosensor
  • S70545: Muscarinic 4 Receptor assay
   

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