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.