Give an example of how a flurogenic assay works to detect the products of an enzymatic...

Give an example of how a flurogenic assay works to detect the products of an enzymatic reaction.

Expert Solution

Enzyme assays are laboratory methods for measuring enzymatic activity.

Enzyme activity = moles of substrate converted per unit time = rate × reaction volume.

when a molecule emits light of one wavelength after absorbing light of a different wavelength, is called flouescence. Fluorometric assays use a difference in the fluorescence of substrate from product to measure the enzyme reaction. These assays are in general much more sensitive than spectrophotometric assays, but can suffer from interference caused by impurities and the instability of many fluorescent compounds when exposed to light.

An example of these assays is the use of the nucleotide coenzymes NADH and NADPH. Here, the reduced forms are fluorescent and the oxidised forms non-fluorescent. Oxidation reactions can therefore be followed by a decrease in fluorescence and reduction reactions by an increase. Synthetic substrates that release a fluorescent dye in an enzyme-catalyzed reaction are also available, such as 4-methylumbelliferyl-β-D-galactoside for assaying β-galactosidase.

queen_honey_blossom answered 1 year ago

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