The precipitate collected from step A was than dissolved in 1M Tris HCl buffer and dialyzed and these samples
were use for further purification steps.
The samples obtained by dialysis were dissolved in 0.05M Tris HCl (pH 8.4) buffer and loaded on column preequilibrated
with 0.05M Tris HCl Sephadex G 50. It was eluted with 0.05M Tris HCl (pH 8.4) buffer also containing
0.1M KCl, and fractions were collected at the flow rate of 50ml/25min. Fraction viewing high activity were pooled
and asparaginase was assayed by previously described assay method.