Isolation and Purification of Human Platelet Arachidonate 12-Lipoxygenase
Klaeren, Stephen A.
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Arachidonic acid 12-lipoxygenase was isolated from lysed human platelets and found to be very stable at temperatures near O°C. Ammonium sulfate fractionation was found to be a useful preliminary purification step giving a 2.4 fold purification. The enzyme eluted off a DEAE-Sephadex A-50 column in 2 separable fractions. Radioimmunoassay and radio thin-layer chromatography was found to be the only reliable method of measuring low level enzyme activity, spectrophotometric methods not being nearly sensitive enough. Evidence was seen that there may be a cofactor associated with the enzyme which is necessary for lipoxygenase activity.