FIG. 5.

SDS-PAGE used for monitoring purification of AdpB and Southwestern blotting with the active fraction after heparin affinity column chromatography. Lanes M to 6, SDS-PAGE; lanes 7 and 8, native PAGE. The molecular size markers in lane M were phosphorylase b (94 kDa), bovine serum albumin (67 kDa), ovalbumin (43 kDa), carbonic anhydrase (30 kDa), and trypsin inhibitor (20 kDa). Lane 1, the cell lysate from S. griseus HH1 (1 mg of protein); lane 2, after ammonium sulfate fractionation (1 mg of protein); lane 3, after DEAE-Toyopearl chromatography (0.5 mg of protein); lane 4, after Mono Q chromatography (0.5 mg of protein); lane 5, after heparin affinity chromatography (0.2 mg of protein); lane 6, after elution from nondenaturing polyacrylamide gel (0.1 mg of protein); lane 7, nondenaturing gel electrophoresis of the active fraction after heparin affinity chromatography; lane 8, autoradiogram of the same sample as in lane 7 subjected to Southwestern blotting with the ³²P-labeled EcoRI-Eco47III fragment.