[CH3:1][CH2:2][CH2:3][CH2:4][CH2:5]/[CH:6]=[CH:7]\[CH2:8]/[CH:9]=[CH:10]\[CH:11]=[CH:12]\[CH:13]=[CH:14]\[C@@H:15]1[O:17][C@H:16]1[CH2:18][CH2:19][CH2:20][C:21]([OH:23])=[O:22].CCCCC/C=C\C/C=C\C=C\C=C\[C@@H]1[O:40][C@H]1CCCC(OC)=O.[Na+].[Cl-].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O>P([O-])([O-])([O-])=O.[K+].[K+].[K+]>[CH3:1][CH2:2][CH2:3][CH2:4][CH2:5]/[CH:6]=[CH:7]\[CH2:8][C@@H:9]([OH:40])/[CH:10]=[CH:11]/[CH:12]=[CH:13]/[CH:14]=[CH:15]\[C@@H:16]([OH:17])[CH2:18][CH2:19][CH2:20][C:21]([OH:23])=[O:22]The LTA4 substrate was freshly prepared by alkaline hydrolysis of LTA4 methyl ester (Cayman Chemical Co., USA). After incubating for 10 minutes in the presence of LTA4 (1 μM in 50 mM Tris-HCl, 0.15 M NaCl, 0.5% BSA, pH 7.4), the reaction is stopped by diluting 1/20 in 0.1 M potassium phosphate buffer containing 1.5 mM NaN2, 0.4 M NaCl, 1 mM EDTA, 0.1% BSA, pH 7.4, −4° C.
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