Brief Definition of ELISA Terms

ELISA TermDefinition
Solid phase
Usually a microtiter plate well. Specially prepared ELISA plates are commercially available. These have an 8 × 12 well format and can be used with a wide variety of specialized equipment designed for rapid manipulation of samples including multichannel pipets.
The process of adding an antigen or antibody, diluted in buffer, so that it attaches passively to the solid phase on incubation. This is a simple way for immobilization of one of the reactants in the ELISA and one of the main reasons for its success.
Washing The simple flooding and emptying of the wells with a buffered solution to separate bound (reacted) from unbound (unreacted) reagents in the ELISA. Again, this is a key element to the successful exploitation of the ELISA.
Antigens A protein or carbohydrate that when injected into animals elicits the production of antibodies. Such antibodies can react specifically with the antigen used and therefore can be used to detect that antigen.
Antibodies Produced in response to antigenic stimuli. These are mainly protein in nature. In turn, antibodies are antigenic.
Antispecies antibodies Produced when proteins (including antibodies) from one species are injected into another species. Thus, guinea pig serum injected into a rabbit elicits the production of rabbit anti¨Cguinea pig antibodies.
Enzyme A substance that can react at low concentration as a catalyst to promote a specific reaction. Several specific enzymes are commonly used in ELISA with their specific substrates.
Enzyme conjugate An enzyme that is attached irreversibly to a protein, usually an antibody. Thus, an example of antispecies enzyme conjugate is rabbit antiguinea linked to horseradish peroxidase.
Substrate A chemical compound with which an enzyme reacts specifically. This reaction is used, in some way, to produce a signal that is read as a color reaction (directly as a color change of the substrate or indirectly by its effect on another chemical).
Chromophore A chemical that alters color as a result of an enzyme interaction with substrate.
Stopping The process of stopping the action of an enzyme on a substrate. It has the effect of stopping any further change in color in the ELISA.
Reading Measurement of color produced in the ELISA. This is quantified using special spectrophotometers reading at specific wavelengths for the specific colors obtained with particular enzyme/chromophore systems. Tests can be assessed by eye.