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. 2013 Jan 17;288(9):6726–6742. doi: 10.1074/jbc.M112.444034

FIGURE 1.

FIGURE 1.

Identification of MARCKS and BASP1 as PSA binding partners using a PSA-mimicking scFv antibody for affinity chromatography. A, purified scFv antibody was immobilized and used for affinity chromatography with soluble brain proteins isolated from postnatal (0–7 days old) mice. After collecting the flow-through (FT) and three consecutive wash fractions (W1, W2, W3), proteins bound to the scFv antibody were eluted. The eluate (E), the flow-through, the wash fractions, and the purified scFv antibody (sFv) were separated by SDS-PAGE and subjected to silver staining or Western blot analysis using the scFv antibody. B, the eluate was separated by two-dimensional gel electrophoresis, pH 3–10, and the two-dimensional gel was subjected to silver staining. A and B, the position of the ∼70- and ∼50-kDa protein bands, which were identified by mass spectrometry as MARCKS and BASP1, are indicated by an arrow and arrowhead, respectively. The asterisk indicates the position of the scFv antibody.