Fig. 2. Generation and purification of recombinant and soluble SFRP1. (A) The conditioned media (CM) from MDCK/control or MDCK/SFRP1 mass cultures (MC) and few of the derived clones were tested for the presence of secreted SFRP1 by immunoblot with anti-myc antibody. Clones 2, 3 and 10 secreted the highest yield of SFRP1 compared with the yield from MC or clone 4. (B) Partial purification of SFRP1. Concentrated CM from either clone 10 [see (A)] or MDCK/control were applied to Ni⁺⁺-NTA agarose column and eluted with 100 mM imidazole. Eluted fractions were separated by 10% SDS-PAGE. The protein content of a representative fraction eluted from control (lanes 1 and 3) or clone 10 (lanes 2 and 4) media were compared. The identity of SFRP1 in the fraction was confirmed by immunoblot with anti-myc antibody.