Enzymes have an important role in nearly all cellular processes. Therefore, the ability to study the activity of specific enzymes is vital in biological research. Enzyme assays commonly measure either the consumption of the substrate or the rate at which product is produced over time. End point assays can be performed with the use of a stop reagent to prevent any further enzyme activity from occurring. Several methods of measuring enzyme activity are available and the same enzyme can often be assayed in a variety of ways.
Marker Gene Technologies, Inc. provides a wide range of products for your enzyme analyses allowing you to find the perfect solution for your research. These products are divided into 2 categories:
Enzyme substrates are usually a synthetic molecule that mimics the natural substrate of the enzyme of interest but with the addition of a reporter molecule that allows for the detection of enzyme activity. Most commonly this detection occurs by the reporter molecule being in some way activated by the removal of the target molecule of the enzyme leading to an increase in signal. This reporter molecule could be a fluorescent dye that is quenched by the presence of the substrate molecule and upon cleavage becomes fluorescent again, this means that the level of fluorescence is directly related to the amount of enzyme activity. As enzyme reactions are kinetic it is possible to see measurable signal when only a small amount of enzyme is present allowing for highly sensitive assays.
Enzyme inhibitors are molecules that bind to an enzyme and cause a reduction in activity. Enzyme inhibitors generally act in one of two ways, by entering the active site and blocking substrate entry or by binding to the enzyme itself and causing some sort of chemical or conformational change that renders it inactive. Enzyme inhibitors can be classified into two further subtypes: reversible and irreversible inhibitors, the later most often being those that cause conformational changes within the enzyme structure. Reversible inhibitors are often those that bind to the active site weakly and can therefore be displaced or otherwise become unbound to the active site.
Enzyme inhibitors can be a useful tool for research, inhibiting a certain enzyme can show what effects that enzyme has on a particular pathway, or inhibiting an enzyme within a live cell environment can lead to a useful disease model for studying potential therapeutics.