Skip to main content
Figure 2 | Lipids in Health and Disease

Figure 2

From: Purification and biochemical characterization of a secreted group IIA chicken intestinal phospholipase A2

Figure 2

Purification of ChPLA2-IIA. (A) Gel filtration chromatography of intestinal ChPLA2-IIA on Sephadex G-50. The column (1.5 cm × 34 cm) equilibrated in 20 mM Tris-HCl buffer pH 8.0 containing 20 mM CaCl2 and 2 mM benzamidine. Elution was performed with the same buffer at a flow rate of 40 ml.h-1 and 3 ml samples were collected. ChPLA2-IIA activity was measured as described in Material and methods section using egg yolk emulsion as substrate. The pooled fractions containing the PLA2 activity were indicated by horizontal line. (B) Mono-S Sepharose chromatography. The column (5 cm × 2 cm) was equilibrated with 20 mM Tris HCl buffer pH 8.0 containing 20 mM CaCl2 and 2 mM benzamidine; and then washed with the same buffer containing 0.3 M NaCl. Linear salt gradient (0.3 to 1 M NaCl, dotted line) was applied to the column; gradient chamber 75 ml; 2 ml fraction; flow rate, 40 ml/h. The pooled fractions containing the PLA2 activity were indicated by horizontal line. SDS-PAGE (15%) analysis of pure ChPLA2-IIA was inserted in Figure 2B. Lane 1, molecular mass markers (MM); Lane 2, 15 μg of proteins obtained after Sephadex G-50 chromatography; Lane 3, 15 μg of purified ChPLA2-IIA, obtained after Mono-S chromatography.

Back to article page