Rotation of plates while incubating reagents
- In certain ELISA systems, the plates are rotated during incubation for better antigenantibody reaction.
- The effect of rotating plates is to mix the reactants completely during the incubation step.
- Since the solid-phase limits the surface area of the absorbed reactant, the mixing ensures that, potentially reactive molecules are continuously coming into contact with the solid-phase.
- During stationary incubation, mixing only takes place because of diffusion of reagents.
- Thus, to allow maximum reaction from reagents in stationary conditions, greater times of incubation may be required, than if they are rotated.
- Rotation also allow ELISA to be performed independent of temperature conditions.
- The interaction of antigen & antibodies relies on their closeness, and the kinetic energy provided to the system, which is encouraged with the mixing during rotation.
- Stationary incubation relies on the diffusion of molecules & thus is dependent on temperature.