“…After 16 hours the pituitary glands were removed and immediately placed in ice cold 145 mM NaCl, 50 Mm Na2HPO4, 0-1% Na,S203, pH 7.2 (phosphate buffered saline with sodium thiosulphide, PBST), homogenised with a loose fitting glass homogeniser, and centrifuged at 27 000 x g for 10 minutes. The clear supernatant was run in a preparative isoelectrofocusing column with a linear sucrose gradient.20 Ampholine (LKB), pH 3-5-10 or 4-6, was used as the carrier electrolyte; the anode solution was 0 1 M glutamic acid, 0 5 M phosphoric acid and the cathode solution Fractions were asayed for pH and dialysed against PBST for 48 hours and then tested for antisecretory activity.…”