ABSTRACT
One key consideration during the enrichment of basophils is preventing their stimulation and degranulation. The method described is based on the report of Leonard et al.
• Venous blood, freshly drawn • 0.1
M
EDTA solution, pH 7.4 • Percoll gradients. Prepare a Percoll (Amersham Pharmacia Biotech)
stock solution by mixing 90 ml Percoll, 9 ml 10
×
HBSS, 1 ml 0.25
M
HEPES buffer pH 7.3, and 0.4 ml 1
M
HCl. Dilute with 1
×
HBSS to prepare solutions of 1.070 g/ml (24 ml Percoll stock + 20 ml HBSS); 1.079 g/ml (27 ml Percoll stock + 15.9 ml HBSS); and 1.088 g/ml (23 ml Percoll stock + 10 ml HBSS).
