Human Glucagon-Like Peptide-1 Receptor Reporter Assay System

This assay utilizes proprietary human cells that provide constitutive expression of the Human Glucagon-like Peptide-1 Receptor (GLP-1R). The GLP-1R belongs to a class B G protein-coupled Receptor (GPCR), which is activated by several forms of GLP-1. GLP-1 is the second incretin hormone identified in the intestinal epithelial endocrine L-cells, followed by gastric inhibitory polypeptide (GIP). GLP-1/GLP-1R mainly regulates insulin secretion in response to high blood glucose levels. This receptor system plays a crucial role in energy homeostasis. Upon the ligand binding of GLP-1, GLP-1R signals through Gαs to drive an increase in intracellular cAMP via activation of adenylate cyclase (AC). This signaling pathway continues through the activation of protein kinase A (PKA) and exchange protein activated cAMP (EPAC) dependent mechanisms. Ultimately, the signal transduction cascade stimulates the opening of calcium and cation channels to induce calcium influx, which promotes insulin secretion. cAMP response binding element (CREB) is also activated by GLP-1R to induce the expression of insulin transcription factor in cAMP/PKA-dependent manner.

INDIGO's Reporter Cells contains an engineered luciferase reporter gene functionally linked to tandem Cyclic AMP Response Elements (CRE) and a minimal promoter. Activated adenylate cyclase results in the production of cAMP, which binds the transcription factor CREB (cAMP Response Element-Binding Protein). Activated CREB binds to CRE sequences, seeding the formation of a complete transcription complex that drives luciferase gene expression. Quantifying relative changes in luciferase enzyme activity in the treated reporter cells relative to the untreated reporter cells provides a sensitive surrogate measure of drug-induced changes in GLP-1R activity. Accordingly, the principal application of this reporter assay is in the screening of test compounds to quantify any functional activities, either activating or inhibitory, that they may exert against GLP-1R.

INDIGO’s Reporter Cells are transiently transfected and prepared as cryopreserved stocks using a proprietary CryoMite™ process. This cryo-preservation method allows for the immediate dispensing of healthy, division-competent reporter cells into assay plates. There is no need for intermediate treatment steps such as spin-and-rinse of cells, viability determinations or cell titer adjustments prior to assay setup.

This assay kit provides the convenience of an all-inclusive cell-based assay system. In addition to GLP-1R Reporter Cells, provided are two optimized media for use in recovering the cryopreserved cells and for diluting test samples, the reference activator GLP-1, Luciferase Detection Reagents, and a cell culture-ready assay plate. [INDIGO Catalog Nos. IB33001 | IB33002]

WARNING This product is not for human or veterinary use.