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Reporter gene assays for investigating GPCR signaling.

Methods Cell Biol.. 2017; 
Azimzadeh P, Olson JA Jr, Balenga N.
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Peptide Synthesis ... 1. CaCl 2 as 1 M stock in distilled water. Store at 4°C. 2. SrCl 2 as 1 M stock in distilled water. Store at − 20°C. 3. Human P-15 peptide (Balenga et al., 2017)(GenScript): Dissolve in DMSO at 10 mM concentration. Store at − 80°C. 4. ... Get A Quote

摘要

Luciferase-based assays are applied to evaluate various cellular processes due to their sensitivity and feasibility. The field of GPCR research has also benefited from this enzymatic reaction both in deorphanization campaigns and in delineation of the signaling pathways. Here, we describe the details of this assay in GPCR studies in 96-well format and will provide examples where the assay can show constitutive activity of an orphan GPCR and demonstrate the impact of cell type on the efficacy and potency of ligands.

关键词

Adhesion GPCR; Biased signaling; CaSR; Luciferase; Transcription factors