Assay Method Information

Assay Name:  In-vitro LOX Inhibition Assay
Description:  It is determined by kinetic mode of spectro-photometric determination method,which was performed by recording the rate of change in absorbance at 234 nm. It is based on the principle that the increase of absorbance at 234 nm due to formation of 9- and 13- hydroperoxides by LOX with substrate (linoleic acid). This reaction does not occur as the enzyme gets inhibited due to addition of inhibitor in it. Briefly, the stock solution of lipoxygenase was prepared by dissolving 10,000 units/ml of Lipoxidase in 200 mM borate buffer, pH 9, and 5 uL of 80 mmol linoleic acid. Six different concentrations of inhibitors were selected, and made for determining the enzyme inhibition activity. Increasing concentrations of inhibitors were added to enzyme solution and kept for 5 min followed by substrate addition and kept at room temperature with stirring for 20 min. One sample was also kept which does not contain any inhibitor instead it contain vehicle of inhibitor solution.
Affinity data for this assay
 

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