A novel HepG2/tGFP-S1P1 cell line has been developed through stable transfection to modulate the internalization of the bHuman Sphingosine-1-phosphate receptor 1 (S1P1R) complex in living cells and to assay for compounds inducing tGFP-S1P1R distribution inside the cells. Fluorescent S1P1R cell line was obtained by transfection of an expression vector for a fusion protein of tGFP and human S1P1 Receptor, as well as G418 into HepG2 cells. Single cells with strong green fluorescence were selected by flow cytometry, and allowed to expand. These cells constitutively express the tGFP-S1P1R fusion protein.Assay Details: Innoprot S1P1R redistribution Assay Cell Line has been designed to assay compounds or analyze stimuli for their ability to modulate S1P1 receptor activation and the following redistribution process inside the cells. This highly reproducible assay allows monitoring S1P1 receptor activation and redistribution process in High Content Analysis and fluorescence microscope applications. ThIs internalization assay was validated with an average Z prime of 0.71+/- 0.02 for High Content Screening.
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