I work with a-amylase, pepsin and pancreatin and I have a powder stock stored at -20oC.
Can I prepare enzyme solutions and store them in the freezer and how? Do I need to add glycerol or just MQ water is sufficient?
I have noticed that the a-amylase solution stored at 4oC caused the enzyme inactivation.
Hi Dora,
Temperature ~ 4 degrees C
Buffer solution: appropriate for each enzyme case
Pepsin and pancreatin are proteases, for them you need to use an inhibitor
Check some protocols and a previous discussion:
https://animalscience.unl.edu/Research/RumNut/RumNutLab/07-BasicRulesforHandlinganEnzyme.pdf
http://wolfson.huji.ac.il/purification/PDF/StorageProteins/PIERCE_ProteinStorage.pdf
https://www.researchgate.net/post/How_long_may_enzyme_activity_be_preserved_in_storage_at_4_degree
Thank you very much Marius.
The pH of the buffer should be the optimal for the enzymatic activity, right? Or storage conditions are different from the activity conditions?
This is highly dependent on your enzymes and their provenience. A good online database for enzymes info is BRENDA https://www.brenda-enzymes.org/
Good luck!
Marius
Hi thanks for the information already posted here. I have a similar question. I have pancreatin powder from Sigma stored at -20 deg. I would like to prepare pancreatin solution to be used on zebrafish early embryos to loosen their tissues so that tissues of interest can be removed easily from the embryo.
In the lab we use L15 medium for preparing zebrafish explants. Any ideas on whether I can directly dissolve pancreatin in L15? The assay itself, where embryos will be exposed to pancreatin is for less than 10 minutes.
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