I am going to do in-vitro simulations of the human digestion process, so I am planning to include all the relevant enzymes.
My question is, are the enzymatic assays required after I buy the enzymes from the providers, in order to determine their specific activity? And also, do I need to run these assays every time before the experiments or only once in the beginning is enough? I don't know if the special activity of the enzymes, which come in a lyophilized powder, is affected through time of storage.
Hi there,
Yes it is necessary to check enzyme activity for different reasons:
first you reconstitute enzyme stock solution from lyophilized powder then you have to validate this process by showing the reconstituted enzyme is active and exhibit the expected specific activity,
second you are about to use these enzymes as a mixture then individual activity measurements prove the whole bunch is active at first,
third you have to prove/follow enzyme stability with time if you have to repeat the experiments.
yes, you have to do confirm the specific activity of the enzyme every time , when one has to do the enzyme activity studies. This study will give us the position of protein/enzyme stability and its reproducible functional activity (KM/Vmax) . This is more essential when the enzyme is in solution ( due to pH ) and when enzyme is in lyophilized physiological state (to free from protein decomposition due to moisture/humidity and sometimes photo decay)
You do need to check the activity of the Enzyme everytime before using it in your study. The supplier you obtained the enzyme from should have storage stability information, in terms of lyophilized and even reconstituted conditions, so you have some assurance that you are purchasing the most stabilized enzyme for your work. I am a Principal Scientist at a company that produces a variety of enzymes and we provide the customer with all the detail they need, whether it is published on the website or a quick call to Tech Service to connect the customer to Scientist like myself, to get direct answers.
- Badges
- Science topic
- Similar topics
- Engineering
- Materials Engineering
- Powders