was added to the supernatant. His Bind Column was equi- librated with 1 binding buffer and the solubilized protein was loaded on it. The entire volume was allowed to flow through col-
umn with a flow rate of 100 ll/min. The column was washed with
1 Wash Buffer (4 M NaCl, 480 mM imidazole, 160 mM Tris-HCl, pH 7.9). Finally, protein was eluted with 1 Elution buffer (4 M imidazole, 2 M NaCl, 80 mM Tris-HCl, pH 7.9). The purified protein was analyzed by SDS PAGE (Laemmli, 1970) and stored at 20 C for further use