2.4 Basophil activation test with blood from cat-allergic and non-allergic subjects
Whole blood samples were collected in EDTA tubes no more than 18 hours prior testing. Degranulation of basophils was determined using the FLOW CAST Basophil Activation Test (Bühlmann Laboratories, Schönenbuch, Switzerland) following the producer’s instructions.
Samples from non-allergic subjects were primed with 1 µg/ml IgE for 1.5 hours at 30°C followed by centrifugation at 400xg for 5 min. The supernatant was discarded to remove the unbound IgE Abs, with the cells subsequently washed twice and resuspended in stimulation buffer following the producer’s instructions of the FLOW CAST protocol.