Figure 3.

Secreted proBDNF is dimeric. ProBDNF (non-His-tag) from recombinant adenovirus-infected cell media was purified using cation-exchange chromatography as described. Purified proBDNF was fractionated by Superdex 200 gel filtration in 25 mm Tris-Cl, pH 7.0, and 200 mm NaCl. The indicated fractions were analyzed by Western blotting with anti-BDNF antiserum. Top, Chromatogram of proBDNF elution profile. Native molecular weight standards are indicated by arrows. The peak of eluted proBDNF (arrow) has a calculated molecular weight of 68 kDa. Bottom, Western blot (WB) of eluates containing proBDNF.