In the Indirect ELISA ,
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The ELISA (enzyme-linked immunosorbent assay) is a test which uses color change and antibodies to identify a substance. An indirect ELISA is one of three ELISA tests.
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Direct ELISAs are sometimes refered to as sandwich ELISAs because unlike the indirect ELISA in which the antigen is binded nonspecifically to the ELISA plate, an antibody is first plated that will capture the antigen. Next, an enzyme-linked antibody is plated and lastly a substrate which creates a measurable color change (OD). The two antibodies "sandwich" the antigen.
There is just one enzyme used in the ELISA reaction. This enzyme is linked to the secondary antibody. Commonly used ELISA enzymes are:Alkaline phosphataseHorseradish peroxidase
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enzyme linked immunosorbant assay