Figure 2 | Scientific Reports

Figure 2

From: Fetal bovine serum albumin inhibits antimicrobial peptide activity and binds drug only in complex with α1-antitrypsin

Figure 2

Purification of serum components responsible for AMP-binding. (a) Proteins bound to TP4-A12I,A15I peptides. The components in crude bovine serum were pulled down by biotinylated TP4-A12I,A15I peptides which were immobilized on Streptavidin-conjugated beads, and analyzed by 15% reduced SDS-PAGE. (b) Band shift of TP4-A12I,A15I peptides by serum proteins after ammonium sulfate fractionation. (c) Pulling down of serum proteins after ammonium sulfate fractionation by biotinylated TP4-A12I,A15I peptides which were immobilized on Streptavidin-conjugated beads. (d) FPLC chromatography for the purification of serum proteins from 70% saturated ammonium sulfate fractionation. Interested proteins (A, B, C and D) are shown on the profiles of FPLC Mono-Q (left) and S12 (right) column chromatography. (e, f) Homogeneity of purified serum components analyzed by horizontal 8% native PAGE, pH8.8, and vertical 15% reduced SDS-PAGE, respectively. Bio, biotinylated TP4-A12I,A15I; S.t., Streptavidin-conjugated beads; PD, pulling down; AS70, fractionation by 70% ammonia sulfate saturation.

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