Figure 5. Identification of PACA-modified proteins. (A) Scheme illustrating the oxidation of PACA for reacting with cysteine residue in the proteins. (B) Experimental procedure for the identification of PACA-modified proteins. RAW264.7 macrophages were treated with 50 μM PACA for 24 h. The cellular proteins were biotinylated with azido-biotin under chemistry conditions and separated by streptavidin-coated magnetic beads. (C) Detection of PACA-modified proteins. The protein fractions were resolved by SDS-PAGE and stained with Coomassie Brilliant blue R-250, by streptavidin-HRP conjugate or by antibody against PPAR-γ. M, protein marker; C, cell lysates; F, flow-through, W1-W5, washing fraction 1 to 5; E, elution.