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GPCR Assays

Detecting GPCR-Mediated Responses in Living Cells

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Montana Molecular develops single-color genetically encoded biosensors for detection of signal transduction pathways in living cells. Our sensors fluoresce in either green or red, and they can be combined with one another for the simultaneous detection of multiple analytes.

 

  • Detect kinetic Gs, Gi, and Gq mediated responses in living cells
  • Combine multiple assays in the same cell population
  • Express sensors and run assays in disease relevant cell types
  • Detect fluorescence on imaging systems or automated plate readers (Z’>0.8)

Detecting Gs

Detect Gs mediated cAMP fluctuations with the cADDis cAMP assay.

BMG Clariostar Application Note for Gs

Detecting Gi

Our cADDis cAMP sensor can be also used to detect Gi mediated responses.

Request Gi Assay Paper 

Detecting Gq

Detect Gq mediated responses using R-GECO Ca2+, DAG, and PIP2 assays.

BMG Clariostar Application Note for Gq

cADDis cAMP Assay

DAG Assay

PIP2 Assay

R-GECO Ca2+ Assay

Simple Protocol

We strive to create simple protocols with minimal liquid handling. Cell lysis, IBMX, enzymes, and co-factors are not necessary for these assays. Add our sensors to your cells, incubate, add drug, and measure fluorescent changes.

Continuous Multiplex Measurements

Combine our red and green sensors in the same cells to measure multiple analytes simultaneously. Simply add the both sensors to the transduction mix. Measure two signals, examine ligand bias, deorphanize GPCRs, and look for off-target GPCR activation. 

Assays in Primary Cultures or iPSCs

Detect GPCR mediated responses in cells relevant to your disease, drug target, or biology of interest. Our BacMam-packaged sensors have been used in neurons, cardiomyoctes, islets, and many more primary and iPSC derived cells! Alternate promoters or viral vectors are available by request. The BacMam Transducible Cells page lists some publications showing disease-relevant cells transduced with BacMam. Pictured, at right, are the cADDis cAMP Assay in primary striatal neurons and the R-GECO calcium assay in nCardia’s Cor.4u cardiomyocytes.

Beta-Arrestin Assays

Fluorescent beta-arrestin assays to detect arrestin recruitment at specific GPCRs are available as well. They contain the Arrestin sensor, GPCR and relevant GRK in BacMam, positive control agonist, and sodium butyrate. GPCRs include Angiotensin II (AT1R), Vasopressin (V2R), GLP-1, Mu Opioid (OPRM1), and beta2-adrenergic (B2AR) receptors. We are continuing to validate new receptors, if you do not see your receptor of interest please send us an email!

Quantifying bias is easier than you think.  Check out this video from Pharmechanics to learn how.