I have been trying to monitor kinetics of hydrolysis of different substrates with PSA. PSA was purchased from Sigma Aldrich (Cat # 539832). Even after handling PSA with care (on ice, careful pipetting etc), I noticed that the enzyme activity was lost when the tube was reopened two or three times. Did anyone observe this, and I will be thankful if anyone can give any suggestions on keeping the enzyme active for long time.
Hi,
Actually, if you freeze-thaw your protein more than the first time, then the activity of your protein will gradually loss.I will suggest that you aliquot your substrate into tubes that can be used per time.Try and avoid freeze-thawing cycle because it has great impact on the activity of your protein.
I hope this will help you.
Find an article that can also guide you.
All the best!
I agree with Adesola. Proteinases should always be aliquoted and stored at -80oC to prevent autolysis and denaturation from freeze-thaw cycles.
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