[3H]BK Binding Competition Assays

Affinity for the B₂R was evaluated using a radioligand binding competition assay performed at 0°C in the presence of peptidase inhibitors that included captopril and PMSF (Charest-Morin et al., 2014). Briefly, the binding of 3 nM [³H]bradykinin (Perkin Elmer Life Sciences; 90 Ci/mmol) to adherent intact Human Embryonic Kidney (HEK 293) cells stably expressing the myc-tagged rat B₂R construction (Charest-Morin et al., 2017) was applied to construct binding competition curves for a series of unlabeled peptides.