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Assay Method Information

Assay Name:  Determination of 5-LO Product Formation in Cell-Free Systems (IC50).
Description:  For determination of the activity of 5-LO in 100000g supernatants, aliquots of the supernatants were added to reaction mix, and were preincubated with the test compounds before arachidonic acid was added to start 5-LO product formation. Formed 5-LO metabolites were extracted and analyzed by HPLC. 5-LO product includes LTB4 and its all-trans isomers, and 5(S)-hydro(pero)xy-6-trans- 8,11,14-cis-eicosatetraenoic acid. IC50s are expressed as mean +/- SE, obtained from measurements at 4-5 different concentrations of test compounds, and determined using Graphpad Instat (Graphpad Software Inc., San Diego, CA). EC50/IC50 values were obtained from product formation in PMNL intact cells.
Affinity data for this assay
 

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Last update November 1, 2007
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