Classification of Enzymatic Assay Methods

In most cases, the second step in the assays comprises initiation and incubation. A reaction can be initiated by the addition of the enzyme preparation to the substrate in the reaction mixture, or vice versa. This step is considered the start of the reaction, and all subsequent time points are related to this time. 

Many reactions require termination, which is the step that brings about the cessation of catalysis and thus stops the reaction. Termination may be achieved in several ways, usually via inactivation of the enzyme. 

Termination is often followed by separation of the components in the reaction mixture. Most often separation involves isolating the substrate from the reaction product. 

Detection, the fifth step, refers to that process by which the amount of product formed by the enzyme during a specific incubation interval is determined. 

The last step in an assay involves reduction of the data. This step includes all procedures in which the data are analyzed and graphed to determine initial rates as well as kinetic constants. 

---