GLP1R Activation Assay graph

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Human GLP-1R Reporter Assay Kit

SIZE SKU PRICE
1 x-384 well format assays—
1 x-96 well format assays—
SIZE SKU
1 x-384 well format assays
1 x-96 well format assays
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Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Glucagon-Like Peptide-1 Receptor (GLP-1R). INDIGO’s GLP-1R reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the GLP-1R. In addition to GLP-1R Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against GLP-1R. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.

Features

  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeGPCR
SpeciesHuman
Receptor FormHybrid
Assay ModeAgonist, Antagonist
Kit Components
  • Human GLP-1R Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • Glucagon-like Peptide-1, (7-36), (ref. agonist; in PBS/0.1%BSA)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

Activation of GLP-1R. Activation assays were performed using the reference compounds Glucagon-like Peptide-1 (GLP-1 [7-36]; provided), Liraglutide, Exendin-4 (48- 86) amide acetate (all from Cayman Chemical, Ann Arbor, MI), GLP-1 (7-37) (R&D System, Minneapolis, MN), and Danuglipron (Adoq, Irvin, CA). Luminescence was quantified and values of average (n=3) relative light units (RLU), corresponding standard deviation (SD), Fold-Activation, and Z’ values were calculated. GraphPad Prism software was used to plot data using the least-squares method of non-linear regression for Fold-Activation or RLU vs. Log10 [Cmpd, nM], and to determine EC50 / IC50 values.
Inhibition of GLP-1R. GLP-1R reporter cells were co-treated with an EC80 concentration of the reference activator GLP-1 (7-36) and varying concentrations of the GLP1R specific inhibitor VU 0650991 (R&D System, Minneapolis, MN). INDIGO’s Live Cell Multiplex (LCM) Assay confirmed that no treatment concentrations were cytotoxic (data not shown). Luminescence was quantified and values of average (n=3) relative light units (RLU), corresponding standard deviation (SD), Fold-Activation, and Z’ values were calculated. GraphPad Prism software was used to plot data using the least-squares method of non-linear regression for Fold-Activation or RLU vs. Log10 [Cmpd, nM], and to determine EC50 / IC50 values.

Target Background

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 GLP1, GLP1R 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.

Because of the significant role of GLP-1/GLP-1R as a key regulator of metabolism, several clinical trials have been attempted to develop therapies for patients with Type II diabetes. For example, GLP-1R agonists (GLP-1RAs) have been introduced as a novel class of therapeutic agent to manage glycemic control.

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