What's the purpose of using an enzyme-linked to anti-IgG in the ELISA test?

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  • Anonymous
    2 months ago

    Most ELISAs use a detecting antibody which is normally of the IgG class.  Such as a mouse anti-T4 antibody to detect T4 in some samples.  The antibody was made in a mouse.  The anti-mouse secondary with the linked enzyme is generally used for two reasons, one of them assay-development related and the other related to assay performance.  Linking the enzyme to the antibody is not 100% efficient, and requires the 'wasting' of some of the antibody.  Antibody production for specific molecules of interest results in antibodies that may have limited availability, and so wasting a bunch of it to get the enzyme tag is not prudent.  The other reason is that using a secondary antibody allows for amplification of the signal, and is another parameter of the assay, which can lead to more fine tuning as far as concentrations, etc.

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