Abstract:This unit describes the over-expression and purification of active serine proteases and their variants from E. coli inclusion bodies. The strategy includes the folding and purification of a stable zymogen precursor protein, and its later activation with the appropriate convertase to the less stable but active protease. A test to follow the presence of activity in the samples, together with an active-site titration protocol to determine the number of active sites per mole of total protein are provided. It shoul… Show more
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