Reporter Functional Assay Services

For drug discovery and development, Creative Biolabs offers a variety of reporter functional assay services to assist you to evaluate the metabolic and safety profiles of lead candidates on a large scale.

Reporter functional assays work by using GPCR-mediated secondary messengers like cAMP or calcium to activate or inhibit a responsive region upstream of a minimal promoter, which then controls the production of a specific reporter protein. β-galactosidase, luciferase, green fluorescent protein, and β-lactamase are examples of often used reporters.

When agonists bind to G_s-coupled GPCRs, intracellular cAMP rises, which activates protein kinase A (PKA), phosphorylating a CRE-binding protein (CREB), and then binds to a CRE in the promoter of a target gene and stimulates transcription. G_q-coupled GPCRs can promote gene expression by inducing intracellular calcium influx, which dephosphorylates cytoplasmic NFAT, activates the NFAT response element, and dephosphorylates cytoplasmic NFAT. Meanwhile, protein kinase C (PKC)-dependent MAP kinase activation via G_q-coupled receptors can trigger SRE-mediated reporter-gene transcription. The stimulation of Gi-coupled GPCRs lowers intracellular cAMP levels and inhibits CRE-mediated reporter gene transcription.

Fig.1. Schematic diagram showing GPCRs signaling pathways. (Chen, 2015)

Luciferase Assay Services

Creative Biolabs offers luciferase assay services to assist you evaluate the lead candidates on a large scale.

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β-galactosidase Assay Services

Creative Biolabs offers β-galactosidase assay services to speed up your GPCR drug candidate screening and profiling.

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Advantages and Disadvantages of Reporter Functional Assays

For GPCR drug discovery, reporter assays are a common and cost-effective high-throughput functional homogenous assay technology. Miniaturization to a 1536-well size and full robotic automation are two advantages of this platform. The requirement for long incubation periods and the fact that the signaling event assessed is remote from receptor activation, which might result in a significant number of false-positives, are also disadvantages of this platform.

Our Feature Service

• Customized GPCR cell lines.
• Multiple assay formats including calcium flux, cAMP determination, reporter gene assays, etc.
• Determine drug potency and efficacy in agonist, antagonist, inverse agonist, and allosteric modulator modes.
• Stringent QC standards with competitive prices.
• Quick turnaround times of 1-3 weeks depending on project types.

Scientists in Creative Biolabs explored and optimized the GPCR assay. We offer a comprehensive platform to speed up your GPCR drug candidate screening and profiling. Please contact us to discuss your project.

Reference

1. Chen, Y., et al. Luciferase reporter gene assay on human 5-HT receptor: which response element should be chosen? Scientific reports. 2015; 5(1): 1-7.