It has been shown by Asada and colleagues that APX activity, in particular chlorplastic one, quickly disappears from tissue extracts if extracts are made in the absence of added ascorbate (e.g. Chen and Asada 1989 Plant Cell Physiol 30: 987-.). I know this phenomenon so i added ascorbate in the reaction buffer but was it neccessary to be added during the extraction when enzymes are very liable to inactivation and degradation? Could for example the differences in APX activity and subsequent, be explained based on a higher breakdown because of less ascorbate present?
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