Assay of cytoplasmic phospholipase A2.

Assay of cytoplasmic phospholipase A2 (cPLA2) using ß-BODIPY® FL C5-HPC (Cat. No. D3803) as a substrate. The substrate was incubated in enzyme-free assay buffer (lane 1), with secreted PLA2 (from Naja mossambica; lane 2) or with purified human recombinant cPLA2 (lane 3). Cleavage products were separated by thin-layer chromatography in chloroform/methanol/acetic acid/water (50:30:8:4) and were subsequently analyzed using a fluorescence image scanner. Both phospholipases liberated fluorescent BODIPY® FL dye–labeled fatty acids (FFA) by cleavage of the substrate at the sn-2 acyl bond. Figure reproduced with permission from J Biol Chem 274, 19338 (1999).

Assay of cytoplasmic phospholipase A<sub>2</sub> (cPLA<sub>2</sub>) using ß-BODIPY® FL C<sub>5</sub>-HPC (Cat. No. D3803) as a substrate. The substrate was incubated in enzyme-free assay buffer (lane 1), with secreted PLA<sub>2</sub> (from <i>Naja mossambica</i>; lane 2) or with purified human recombinant cPLA<sub>2</sub> (lane 3). Cleavage products were separated by thin-layer chromatography in chloroform/methanol/acetic acid/water (50:30:8:4) and were subsequently analyzed using a fluorescence image scanner. Both phospholipases liberated fluorescent BODIPY® FL dye–labeled fatty acids (FFA) by cleavage of the substrate at the <i>sn</i>-2 acyl bond. Figure reproduced with permission from J Biol Chem 274, 19338 (1999).

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