Fig. 1.
Purification and binding of PpMC-179.
(A) Purification of PpMC-179 from culture supernatant. Proteins were separated on a nonreducing SDS-PAGE gel and stained with Coomassie blue. MW indicates molecular weight standards; Ni, protein eluate from Ni-NTA chromatography; and C4, pooled protein fractions from reverse phase C4-HPLC column. Sizes of molecular weight standards are illustrated. (B) Binding of PpMC-179 to CD36. CD36 bound to PVDF membranes was detected using MoAb-179 followed by HRP-conjugated goat antimouse IgG and visualized using chemiluminescence. Proteins bound to Ni-NTA beads are shown and include PpMC-179 purified on a C4 column (final product), PpMC-179 from the Superdex-75 (S-75) column, and CSA-163 as the negative control. Lanes 1 to 4 had 5 μg, 1 μg, 0.5 μg, and 0.05 μg recombinant protein on beads, respectively.