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The plasma membrane-associated lipoxygenase activity of cell lines derived from human breast cancer was quantitated by a colorimetric microplate assay. 125I-labelled arachidonic acid (AA) was incubated with cell homogenates, and the radiolabelled AA metabolites formed were quantitated colorimetrically following solid phase extraction. The fatty acid composition of the metabolites was characterized using argentation thin layer chromatography (TLC). The monohydroxy fatty acids formed were quantitated by radioautography. A positive correlation between the microplate assay and TLC analysis was observed for the samples containing the highest lipoxygenase activity. The data from the TLC analysis for the samples containing the lowest lipoxygenase activity were below the saturation level of the method and were not considered reliable. The colorimetric microplate assay also demonstrated a linear relationship between the lipoxygenase activity and incubation time. The microplate assay was also used to quantify the lipoxygenase activity of cell lines derived from human breast cancer tissue. This method can be used to measure the lipoxygenase activity of rapidly proliferating cells and to quantitate the lipoxygenase activity in tissues with a high content of lipoxygenase.The present invention relates generally to a method of fabricating micro-heaters for heating microfluidic devices, and in particular to methods of fabricating micro-heaters for fabricating microfluidic devices.
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