RP-HPLC column and SDS-PAGE (15%) of SPLA2. (A) RP-HPLC on a eurospher 100, C-8 column, elution was performed at room temperature within 30 min using a gradient from 0 to 80% solvent B at a flow rate of 1 ml/min. [solvent A, water/trifluoroacetic acid (1000:1, v/v) and solvent B, acetonitrile]. The effluent was monitored at 280 nm. The gradient is indicated by the dotted line. AU: Arbitrary Units. (B) SDS-PAGE (15%) of pure SPLA2. Lane1, molecular mass markers (Pharmacia); lane 2, 7 μg of SPLA2 eluted from RP-HPLC. The gel was stained with Comassie blue to reveal proteins.