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Figure 6


Fig. 6. Purification process and enzymatic activity of ADAMTS12. (A) Western blot analysis of ADAMTS12 during the purification process. c-EBNA and EBNA-TS12 indicate cell-layer-associated fractions from 293-EBNA cells transfected with an empty vector, and the equivalent fraction from 293-EBNA cells expressing ADAMTS12, respectively. Recombinant purified ADAMTS12 is also indicated. (B) Following incubation of aggrecan with recombinant ADAMTS4 (lane 2), purified ADAMTS12 (lane 3) or an equivalent fraction purified from c-EBNA cells (lane 1), samples were deglycosylated and the aggrecan degradation products were detected by using the BC-3 antibody. In lane 4, purified ADAMTS12 was previously incubated with the metalloproteinase inhibitor ilomastat (+Ilo). Molecular mass markers (kDa) in both panels are indicated on the left.





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