Figure 5.

Acetic acid buffer pH 6 improves the viability of basophils but is ineffective to strip off IgE from the basophils. Basophils were incubated on ice for 5 min with ice-cold acetic acid buffer pH 6 (AA pH 6). Cells were then washed and cultured in serum-free X-VIVO 15 medium (0.1 × 10⁶ cells/well per 200 µL) in 96-well U-bottomed plate along with IL-3 (100 ng/0.5 million cells) for 24 h. (A) The forward and side scatter plot of AA pH 6-treated basophils following 24-h culture in IL-3 (left panel) and the yield of basophils (mean ± SD, n = 2 donors) (right panel). The values were calculated based on the percentage of cells in the P1 gate of forward and side scatter plot. (B) Efficacy of stripping of basophil surface-bound IgE by AA pH 6 as analyzed by surface staining of IgE and analyses by flow cytometry.