Phospholipase a2 from porcine pancreas

Resorufin, Resorufin sodium salt, naphthofluorescein, palmitoyl chloride, phospholipase A₂ from porcine pancreas, arachidonyl trifluoromethyl ketone, bromoenol lactone, ionomycin, and H₂O₂ 30% (w/w) were purchased from Sigma-Aldrich. DDAO, arachidonoyl chloride, and LY311727 were purchased from Santa Cruz Biotechnology. 7-HCA was purchased from Cayman Chemical. Syto 9 was purchased from Life Technologies. Egg phosphatidylcholine was purchased from Avanti Polar Lipids, Inc.

OptiPrep® (iodixanol 60% (w/v)), CuSO₄, lipoxidase V from Glycine max (EC 1.13.11.12), phospholipase A₂ from porcine pancreas (EC 3.1.1.4), CaCl₂, ascorbic acid, α-tocopherol (≥ 96%), and barbital buffer were from Sigma Aldrich (St. Louis, MO, USA). 5,5-Dimethyl-1-pyrroline N-oxide (DMPO) was from Dojindo (Kumamoto, Japan). Sephacryl S200 was from GE Healthcare Lifescience (Pittsburg, PA, USA). Agarose and BODIPY^581/591 C11 were from Invitrogen (Grand Island, NY, USA). The BCA protein assay was from Pierce (Rockford, IL, USA). Chelex C-100 and Commassie Blue staining solution were from BioRad (Hercules, CA, USA). Phosphate and Tris buffers were prepared and Chelex-treated for removal of contaminant trace metal ions.