Sephadex G-100 gel filtration
The dialysate was subjected to gel filtration on a Sephadex G-
100 column (2.6 cm 150 cm) previously equilibrated with buffer
B (25 mM Tris–HCl, pH 8.0, containing 0.5‰ Triton X-100) and
eluted with the same buffer at a flow rate of 30 mL/h. Protein content
(absorbance at 280 nm) and proteolytic activity were determined.
Fractions showing protease activity were pooled.