ELISA Tips-Incubation

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.


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