The assay was performed as previously described [20] (link). Cells were dual labelled with SNAP-Lumi4-Tb (40 nM) and 500 nM SNAP-Surface-649 (New England Biolabs, Hitchin, UK) for 30 min at 37 °C, in complete medium. After washing, labelled cells were resuspended in HBSS. TR-FRET signals at baseline and serially after agonist addition were recorded at 37 °C using a Spectramax i3x plate reader with HTRF module. GLP-1R clustering was quantified as the ratio of the fluorescence signal at 665 nm to that at 616 nm.
Free full text: Click here
