A novel U2OS/tGFP-PAC1 cell line has been developed through stable transfection to modulate the internalization of the Human Pituitary adenylate cyclase-activating polypeptide type I receptor (PAC1) complex in living cells and to assay for compounds inducing tGFP-PAC1 redistribution inside the cells. Fluorescent PAC1 Internalization A assay cell line was obtained by stable transfection of an expression vector for a fusion protein of tGFP and human Pituitary adenylate cyclase-activating polypeptide type I receptor, as well as G418 into U2OS cells. Single cells with strong green fluorescence were selected by flow cytometry, and allowed to expand. These cells constitutively express the tGFP-PAC1 fusion protein.Assay Details: Innoprot PAC1 Internalization Cell Line has been designed to assay compounds or analyze stimuli for their ability to modulate Pituitary adenylate cyclase-activating polypeptide type I receptor activation and the following redistribution proce Beta inside the cells. This highly reproducible assay allows monitoring PAC1 receptor activation and redistribution process in High Content Analysis and fluorescence microscope applications. This internalization assay was validated with an average of Z=0.84 +/- 0.02 for High Content Screening.