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| Term | enzyme linked immunosorbent assay | ID (Ontology) | MI:0411 (Molecular Interactions) | |||||
| Definition | Following non-covalent binding of a purified primary ligand to a solid phase, a blocking reagent is added to prevent any non-specific binding. A specific antigen is then allowed to bind to the primary ligand. Unbound antigen is removed by washing and a secondary antibody conjugated to an enzyme (e.g. horseradish peroxidase) is added. Following a washing step to remove unbound secondary ligand, the extent to which a chromogenic substrate (e.g. 3,3', 5,5' tetramethyl benzidine chromogen [TMB]) is converted to a soluble coloured product by the conjugated enzyme in a given time is determined by spectrophotometry using a standard microplate absorbance reader. A similar type of approach can be utilized to detect enzymatic activities. The substrate, attached to a solid phase is incubated in the presence of the enzyme and the enzymatic modification is monitored by an antibody that is specific for the modified substrate (for instance a phosphorylated protein).[ PubMed:11906746 ] | |||||||
| Also Known As | "ELISA" ; "elisa" | |||||||
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affinity technology |__solid phase assay____________________ experimental participant identification | |__identification by antibody___________| enzyme linked immunosorbent assay 144 rec. |
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identification by antibody solid phase assay |
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