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Irrespective of plant material (i.e. part or entire plant), we homogenize it as quickly as possible at low temperature (below 4oC). We often prefer carefully using few ml of liquid nitrogen for rapid and uniform homogenization of plant material in extraction buffer. Subsequent to centrifugation at 15,000 xg at 4oC for 15 min, we take supernatant for various enzyme assays (including SOD and well as APX). Regarding how long you need to incubate depends on the activity of enzyme. For SOD, we generally incubate for 15 min to 30 min (one can increase incubation period or if necessary dilute enzyme with buffer). But, for APX we start recording the activity (time dependent decrease in OD) soon after adding substrate. We generally prefer performing these activities at room temperature.

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