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2 concentrations in the medium were determined in triplicate using a commercially available enzyme immunoabsorbent assay (EIA) kit (Cayman Chemical Company, Ann Arbor, MI), following the manufacturer's protocol. The assay was performed in a total volume of 150 μL, with the following components added in 50 μL volumes: standards or biological samples, enzymatic tracer, and specific antiserum. After overnight incubation at 4 °C, the plates were washed, and 200 μL of Ellman's reagent was added to each well. After 1-3 h, the absorbance of each well was measured at 414 nm. A standard curve with values of 50-0.39 pg/mL was used to evaluate the concentrations
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2 concentrations in the medium were determined in triplicate using a commercially available enzyme immunoabsorbent assay (EIA) kit (Cayman Chemical Company, Ann Arbor, MI), following the manufacturer's protocol. The assay was performed in a total volume of 150 μL, with the following components added in 50 μL volumes: standards or biological samples, enzymatic tracer, and specific antiserum. After overnight incubation at 4 °C, the plates were washed, and 200 μL of Ellman's reagent was added to each well. After 1-3 h, the absorbance of each well was measured at 414 nm. A standard curve with values of 50-0.39 pg/mL was used to evaluate the concentrations
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2 activity was determined by means of a coupled enzyme assay using DL-PC as a substrate and lipoxygenase as a coupling enzyme. The linoleic acid released by phospholipase activity was oxidized by lipoxygenase, giving rise to the corresponding hydroperoxide derivative. The phospholipase activity was followed by measuring the absorbance at 234 nm
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2 activity was determined by means of a coupled enzyme assay using DL-PC as a substrate and lipoxygenase as a coupling enzyme. The linoleic acid released by phospholipase activity was oxidized by lipoxygenase, giving rise to the corresponding hydroperoxide derivative. The phospholipase activity was followed by measuring the absorbance at 234 nm
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