RBL-MRGPRX2 cells (2 × 106 cells) were loaded with Fura-2 acetoxymethyl ester (1 μM) in HEPES buffer containing 0.1% BSA for 30 min in the dark at 37°C, followed by 15 min period for de-esterification in dark at room temperature. Cells were washed and resuspended in HEPES-buffered saline and Ca2+ mobilization was measured for the designated time. Calcium mobilization was determined by measuring the fluorescence ratio between dual excitation wavelengths of 340 and 380 nm, and an emission wavelength of 510 nm using a Hitachi F-2700 Fluorescence Spectrophotometer (29 (link)).
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