Figure 3

Purification of Recombinant ZBD. Panel A: shows the silver staining for the high resolution SDS-PAGE analysis for the purification fraction profile from the P30 molecular sieve chromatography. The 8 M Urea 50 mM Tris pH 7.5 solubilized inclusion bodies (lane S2) was concentrated 10 fold (laneS1) as the start material before loading onto P30 column. 15 μl of the fractions gave OD280 reading were subjected to high resolution SDS-PAGE analysis (lane 10–17). The combined fractions #14-#15 (lane p) were concentrated 10 fold (lane pc). Panel B: shows the silver staining for the high resolution SDS-PAGE analysis for the purification fraction profile from the fractions zinc-chelate chromatography. Combining the partial purified fractions containing ZBD from P30 collumn (lane S) were subjected to the zinc-chelate affinity. The unbound fall-through fractions (lane F1 and F2) and the wash buffer fractions are shown (lane W1 and W2). The bound ZBD was eluted with pH 4.5 buffer and the low pH eluted fractions (lane E3-E10).