What is the reason or importance of diluting an enzyme before an experiment?
I'm writing a report on enzyme activity and characterisation and i need help with understanding this: we were told to report on our results for a 1/10 diluted acid phosphatase. I want to know why we need to dilute the enzyme in the first place and why not use the crude enzyme lysate?
The catalytic activity of a dilute enzyme is, in most cases, directly proportional to the enzyme concentration, as long as certain conditions are met. These are (a) the enzyme concentration is much lower than the substrate concentration so that the substrate binding to the enzyme does not significantly reduce the free substrate concentration, and (b) the reaction is running under so-called "initial rate" conditions before the substrate starts to become depleted and the products start to accumulate. If the enzyme concentration is too high, you will not be able make the measurement while it is still under initial rate conditions. That is why the concentrated enzyme solution is diluted.
If you use crude or soli enzyme in reaction it is too much concentrated so it gives very fastest reaction of desier assay method .
For example, it might use up all the substrate, or the amount of product formed may be too high for the measurement technique. In such a situation, you might not be able to tell the difference between different levels of enzyme activity.
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